High-throughput DNA sequencing laboratories
|
|
- Melvin Bell
- 8 years ago
- Views:
Transcription
1 Journal of Biomolecular Techniques 10: ABRF R AB F Capillary Electrophoresis Platforms for DNA Sequence Analysis Elaine R. Mardis Technology Development, Genome Sequencing Center, Washington University School of Medicine, St. Louis, MO Commercial availability of multicapillary DNA sequencing instruments can significantly impact the existing paradigm in high-throughput DNA sequencing facilities, shifting the ratelimiting step away from the electrophoresis and detection step. These instruments, although novel and poorly understood, represent a significant capital investment.this review informs the reader about the state-of-the-art instruments, although capillary sequencers are in a state of almost constant flux, with improvements emerging rapidly. (J Biomol Tech 1999;10: ) KEY WORDS: multicapillary electrophoresis; automated DNA sequencing. ADDRESS CORRESPONDENCE AND REPRINT REQUESTS TO Elaine R. Mardis, Director,Technology Development, Genome Sequencing Center, Washington University School of Medicine, 4444 Forest Park Boulevard, St. Louis, MO ( emardis@ watson.wustl.edu). High-throughput DNA sequencing laboratories are poised on the brink of a paradigm shift in their workflow and operations. The commercial availability of multicapillary sequencing instruments signals that change is imminent and that a careful evaluation of these platforms, their capabilities, and their limitations is in order. This review considers two commercially available instruments, the MegaBACE 1000 DNA Sequencing System from Amersham Pharmacia/Molecular Dynamics (Sunnyvale, CA) and the 3700 DNA Analyzer from Perkin- Elmer Biosystems (Foster City, CA). Both offer simultaneous electrophoresis and detection of samples on 96 capillaries. The MegaBACE has been available for the past year and the 3700 since January of These instruments are relatively expensive, with the MegaBACE retailing at $200,000 and the 3700 at $300,000. As with any instrument, there are associated consumables, including capillary arrays, separation matrix, buffer, and sequencing reagents. The software for data collection and analysis on both instruments operates in a Microsoft Windows NT environment. ADVANTAGES OF CAPILLARIES VERSUS SLAB GELS Ideally, capillary instruments offer several significant advantages over slab gel scanners. The most obvious advantage of multicapillary systems is the elimination of gel plate assembly and gel casting, because a flowable separation matrix is automatically injected into each capillary just before loading new samples. Some care must be taken to ensure that air bubbles are not introduced into the matrix, because their presence in a capillary causes noise from laser light diffraction and instability in the electrophoretic current. Loading samples onto capillaries also occurs in a straightforward manner that is much less error prone than manual application of samples into slab gel wells, especially at 96-lane density. Sample ions load onto a capillary by a process called electrokinetic injection. A high field voltage is applied between the sample and the anode end of the capillary, causing DNA ions to migrate into the capillary. JOURNAL OF BIOMOLECULAR TECHNIQUES, VOLUME 10, ISSUE 3, SEPTEMBER
2 E. R. MARDIS A major advantage of capillary instruments is the elimination of postrun lane retracking. This is a timeconsuming and error-prone step in slab gel analysis, which scales up with increasing lane number. Capillaries do not require retracking because data collection occurs at previously determined spatial locations corresponding to each capillary in the array. A natural consequence of the elimination of retracking is that the time-consuming transfer of gel image files for offline retracking and analysis is no longer necessary. Only 96 sample files, each around 250 kilobytes, are transferred to a host server or computer for analysis and archiving. This advantage immediately appeals to centers such as ours, where tight instrument loading schedules mean that gel image files from 100 or more sequencers must be transferred across busy local area networks at specific times. Comparatively speaking, capillary instruments require much less hands-on time from technicians than slab gel scanners, although to varying degrees. The PE 3700 allows users to place up to four 96-well microtiter plates on the workspace at one time and then performs the loading, electrophoresis, detection, and data analysis for each plate in succession without user intervention. The MegaBACE requires 10 to 15 minutes of user intervention for sample loading from each 96-well plate, after which electrophoresis and detection are performed automatically. One commonly cited advantage of electrophoresis in capillaries over glass plates is the ability to run high field voltages because of the greater heat dissipation capability from capillaries. In practice, however, our electrophoresis duration is roughly equivalent when comparing the 3700 with the shortest run module on the ABI 377 sequencer (3.3 versus 3.5 hours) and is about 1 hour shorter on the MegaBACE (2.3 hours, a 34% improvement) than on the 377. These times primarily reflect changes that we have made to the manufacturer s recommendations to increase readlength, which is a factor of predominant importance in genomic sequencing projects. We hope future improvements to run matrices and run voltage profiles will allow significantly shorter run times. REVIEW OF THE HARDWARE Detection One fundamental difference between these platforms is the detection schema used by each instrument. The MegaBACE is essentially a commercial version of a multicapillary sequencer devised by Richard Mathies 1,2 and uses his concept of scanning the capillary array at a fixed point with a confocal microscope objective to deliver focused laser light and to collect the fluorescent emissions of laser-excited fragments. In this system, it is imperative that the laser beam is focused at the exact center of each capillary to provide maximum power to the sample bands and that the confocal objective collects emissions from these center pixel positions as well. This precise focusing optimizes sensitivity and signal to noise. In production, we routinely test each instrument for focus every 2 weeks, making adjustments if necessary. Photomultiplier tubes (PMTs) detect the bandpass-filtered fluorescent emissions from sequencing fragment bands. Unlike the MegaBACE, the 3700 uses a sheathflow detector system, including a charge-coupled device (CCD) camera. This instrument is a commercial adaptation of work from Kambara s laboratory in Japan. 3 In sheath-flow detection, the sequencing fragment bands are excited and detected as they elute from the anode end of the capillary into the sheathflow cuvette. The emerging fragment forms a plume that is swept off in a sheath flow of polymer through the laser beam path. The resulting emissions are collected by a spectrograph that uses a reflection grating to disperse the light into its component wavelengths and passes that spectrum onto the surface of the CCD camera. The CCD camera converts the resulting information into digital data that are collected for the run. Because the laser beam enters from the right side of the array, a person can easily detect by eye when the beam is out of alignment with the sheath flow, because the samples at the left of the array exhibit markedly weaker signal than those at the right. Correcting the laser beam entry path on the 3700 requires a trained service technician, whereas an experienced laboratory technician can refocus the Mega- BACE optics. Sample Loading Both capillary platforms offer sample loading by electrokinetic injection, although this is manifested differently in each instrument. In general, electrokinetic injection is a sensitive process that is influenced by the ionic strength of a sample, the amount of template DNA in the sample, and the geometry of the electrode and capillary tip within the sample. The process of electrokinetic injection involves the transfer of charged ions in an electrical field onto the capillary separation matrix. Because only ions transfer in this process, no liquid volume loss occurs from the sample. Ideally, this means that samples can be reloaded if a run is aborted or lost for other reasons. The 3700 loading scheme allows the instrument to achieve unattended operation for analysis of up to 138 JOURNAL OF BIOMOLECULAR TECHNIQUES, VOLUME 10, ISSUE 3, SEPTEMBER 1999
3 CAPILLARY PLATFORMS FOR DNA SEQUENCING four 96-well plates (or one 384-well plate) of reactions. The 3700 has a two-tip gantry-mounted autoloader that transfers 2.5-mL sample aliquots from microtiter plate wells into a multiwell loading bar. Each of the injection ends of the capillary array project into each of the wells on the loading bar, and electrokinetic injection takes place when a voltage is applied to the metal loading bar (acting as the cathode). After injection, the wells of the loading bar are washed out and filled with running buffer, and separation by electrophoresis and data collection begin. Because of the sample transfer process described, the sample aliquots transferred are lost after electrokinetic injection, which means that reloading is limited to the available sample volume remaining in the source plate. Another potential source of volume loss from plates queued on the 3700 workspace for loading is evaporation. The amount of evaporative volume loss can be reduced in a variety of ways. First, we have found that turning off the overhead workspace light source in the 3700 reduces evaporation. Second, plates can be covered with a thin adhesive foil tape after resuspension of samples, and the worker can employ the foilpiercing hardware option available from Perkin-Elmer. With this option, the 3700 foil-piercing tips selectively pierce the foil tape over each well immediately before loading to allow pipette tip access for sample transfer to the load bar. This option appears to be robust for 96-well plates in limited testing, although it adds time to the overall workflow. The MegaBACE requires attended operation to load each plate individually. After placing the sample plate onto the cathode stage and closing the stage door, the plate is raised up to the capillary tips. There is a thin platinum electrode positioned alongside each tip. The injection voltage is applied at these electrodes, and the samples migrate onto the capillary. The plate is replaced by a buffer-filled reservoir during electrophoresis and detection, so the sample plate can be immediately returned to a 20 C freezer and reloaded, if necessary, without sample degradation worries. The loading process requires about 10 minutes to complete. Separation Matrices In capillary systems, an injectable matrix functions to resolve sequencing fragments in the applied electrical field. Predictably, the matrix largely determines the potential resolution limit of the sequencing fragments. It also dictates instrument design and workflow. The currently used matrix for the MegaBACE is linear polyacrylamide (LPA) and for the 3700 is Performance Optimized Polymer 6 (POP6). Each has very different physical characteristics LPA is extremely viscous, and POP6 has the consistency of jelly that directly influence their resolution limit. These matrices also differ in their polymer pore size, a characteristic that may underlie the greater tendency of LPA to clog with template DNA during electrokinetic injection. LPA is packaged into individual 2-mL tubes and is injected into capillaries from within a titanium pressure hull under nitrogen gas pressure. This injection requires that a high-pressure nitrogen tank be attached to the instrument. It also requires a relaxation period before sample injection and electrophoresis so that the polymer can retain its native configuration. Overall, the time required for matrix injection, relaxation, and sample loading is about 50 minutes (30 minutes for the hands-off process of matrix relaxation and prerun). On the 3700, the POP6 matrix is supplied in a 200-mL bottle that nests inside the instrument door. A supply line inside the bottle of POP6 is used to supply a syringe pump that first fills the sheath flow cuvette and then injects POP6 into the capillaries under applied syringe pressure. No relaxation time follows this low-pressure injection, but subsequent flush of the cuvette and loading bar wells to clear excess matrix is required. The overall processing time between runs for these functions to complete is 51 minutes. Chemistry Predictably, Molecular Dynamics and Perkin-Elmer advocate the use of their proprietary labeling chemistries and enzymes for their capillary instruments. Amersham has developed specific sequencing reaction kits for the MegaBACE, with altered deoxynucleotide to dideoxynucleotide ratios to provide improved fragment length distributions from electrokinetic injection. Perkin-Elmer initially supported the use of their existing Big Dye terminator kit, although it appears that a similar deoxynucleotide-dideoxynucleotide optimization may be required. In general, the Amersham kits are based on the Dyenamic Energy Transfer and ThermoSequenase DNA polymerase technologies. These include a Dyenamic ET primer kit and a Dyenamic ET terminator kit. Because this instrument has an operating temperature of 44 C, compressions in dye primer reactions occur. To address this issue, Amersham introduced a ditp-containing Dyenamic ET primer kit. Responsive to the urging of many high-throughput sequencing facilities, Molecular Dynamics now offers a set of bandpass filters that allow the user to detect and analyze the Big Dye labeling chemistry from Perkin-Elmer. JOURNAL OF BIOMOLECULAR TECHNIQUES, VOLUME 10, ISSUE 3, SEPTEMBER
4 E. R. MARDIS Perkin-Elmer has, for the near term, focused their development efforts on Big Dye terminator chemistry, although it is also possible to detect and analyze Big Dye primer reactions on the This limited capability primarily reflects the relatively short time that the instrument has been commercially available, and it is anticipated that additional functionality will be introduced over time. REVIEW OF THE SOFTWARE Both instruments offer software that runs in a Windows NT environment. We have installed the version 1.0 software for each instrument. To improve user interaction in a production environment, we created custom interfaces to facilitate sample sheet import from and data export to our Unix databases. The following observations largely reflect our experiences with and opinions about both software packages. Perkin-Elmer 3700 Although NT-based software represents a radical shift from their tradition of Macintosh-based software, the transition seems smooth from a user standpoint. This is mainly because many of the user interface graphics and features of data collection and analysis from the ABI 377 sequencer have been maintained, including the concepts of editable Run Modules, run status monitoring windows, log files, and spreadsheet-based analysis queuing. This is not to say that the 3700 is without its differences; the main one is that the data collection and analysis software interact directly with an Oracle-based instrument database that is resident on the host computer. The Oracle database stores information about scheduled runs, including the run schedule and plate records, run logs and error logs, preference settings for the Data Collection software, and processed, unanalyzed fluorescence data that have been collected from the CCD. The database is integral to the instrument s operation and must be functional for the instrument to function. Other new features of the software include the Data Extractor program that assembles all relevant information to create analyzed sample files, communicating the data to the AutoAnalyzer program that performs the data analysis and basecalling. Presentation of trace data from individual sample files is virtually identical to the Mac-based software, with the ability to toggle between run information, textual sequence, raw data, and processed data extant. MegaBACE 1000 The data collection and analysis software for the MegaBACE pose challenges, perhaps more so for the experienced PE software user than for the neophyte. Data collection occurs through the Instrument Control Manager (ICM), which guides the user through a predetermined workflow. Preparing the instrument for a run involves stepping through the workflow, during which there are instructions presented to the user at one of two separate liquid crystal diodes (LCDs) on the instrument. These LCDs flank the two stages - cathode and anode -that are accessed through doors at the front of the instrument as a means of supplying necessary items such as tubes of LPA for injection and samples for loading. The run setup requires about 10 to 15 minutes of user interaction time before electrophoresis and data collection are underway. Programs are available for monitoring the run, including a Run Image view that provides display information for each capillary at the top half and displays the electrophoretic profile of four color data from any selected individual capillary on the bottom half. Run status is also a part of this display, including time elapsed, PMT voltages, and run temperature. Another useful display is the Current Monitor window, which displays the current values for each capillary throughout the run. This display is useful because it can give the user a quick view of capillaries that are demonstrating significantly reduced current relative to others. These capillaries may turn out to be routinely problematic and require replacement. Alternatively, reduced current may reflect a clogging event where excessive template has interfered with loading of sequencing fragments onto the matrix. Analysis software for MegaBACE reads uses a spreadsheet interface into which the run data from individual or multiple runs are entered and available for analysis. The user selects a basecaller (a data-processing algorithm), and then the input data from individual runs are analyzed. Electropherograms of the resulting data can be viewed by highlighting individual samples and requesting the electropherogram view from a pull-down menu or by selecting all samples in the spreadsheet and viewing the traces in groups of four. In a production setting, the MegaBACE software has posed problems for technicians in our laboratory because the user interface does not allow straightforward import of the sample sheet or run parameter selection. The postrun data analysis should be automated and not require the manual procedures described. On a positive note, the manufacturer is working aggressively to address these issues and to provide a more tractable user interface. Molecular 140 JOURNAL OF BIOMOLECULAR TECHNIQUES, VOLUME 10, ISSUE 3, SEPTEMBER 1999
5 CAPILLARY PLATFORMS FOR DNA SEQUENCING T ABLE 1 Modifications to Suggested Run Parameters Made for the PE 3700 or MD MegaBACE to Improve Performance Instrument Suggested Parameter Modified Parameter Rationale for Modification C oven and sheath-flow cuvette 37 C oven and sheath-flow cuvette Lowering the run temperature temperature temperature (dye terminator decreases the matrix viscosity chemistry only) and allows better fragment resolution range ,000 sec data collection time 12,000 sec data collection time a Additional data collection time required for slower migration of fragments at 37 C MegaBACE Not applicable Preinject samples at 3 kv for Helps to rid samples of excess 20 sec ionic content that can interfere with electrokinetic injection success MegaBACE Loading from a standard Loading from an aqueous Reduces the incidence of overformamide/edta solution 0.1% hydroxyethyl cellulose injected samples solution MegaBACE Inject samples at 10 kv for 15 sec Inject samples at 2 kv for 60 sec Reduces the incidence of overinjected samples MegaBACE Data collection for 90 min Data collection for 140 min Increases the resolution and at 10 kv at 7 kv readlength for most samples to 750 bases aa consequence of longer data collection time on the 3700 is that the sheath-flow volume must be increased to 13,500 L to provide sufficient amounts of polymer for the sheath flow elution of fragments. Dynamics has made this version 2.0 available to beta test sites, including our laboratory. OUR EXPERIENCE WITH CAPILLARY PLATFORMS Two facts regarding our experiences with the MegaBACE and the PE 3700 are important. First, we have had access to MegaBACE sequencers for 2 years and to the 3700 for 5 months. As such, our recent experiences with the 3700 in some ways reflect struggles that we had with the initial MegaBACE units of 2 years past. Second, we address new instrumentation in light of our current workflow and protocols, trying to deviate as little as possible from proven processes and not always considering optional approaches that might be appropriate in another setting. In other words, what is right for our highthroughput sequencing laboratory will not necessarily be right for another and almost certainly will not be appropriate for a lower-throughput or core facility. Both types of capillary instruments have been run at our facility in mock production mode, involving maximizing the throughput per machine per day. We are only now poised to introduce them into our genomic and EST production groups. Running mock production allows us to assess instrument performance at maximum throughput, thereby determining whether there are hardware stability problems. It allows us to gauge whether the preps and sequencing protocols and the run parameters we have optimized are sufficiently robust. Mock production generates feedback from a small user group on what additional hardware and software pieces of the puzzle are still required for straightforward implementation. Modifications Table 1 provides a summary of the modifications we made to existing protocols and procedures associated with each instrument relative to their manufacturer s recommendations. In most cases, these modifications were meant to address shortcomings in the JOURNAL OF BIOMOLECULAR TECHNIQUES, VOLUME 10, ISSUE 3, SEPTEMBER
6 E. R. MARDIS performance of each instrument. In the case of the 3700, we were primarily concerned with short readlengths caused by the low viscosity of the POP6 matrix and to the detection bias of shorter fragment lengths resulting from the mismatch of sheath flow velocity to electrophoretic velocity. For the Mega- BACE, we were concerned with the relatively high percentage of overinjections experienced with the LPA matrix when excess DNA is injected, impeding the flow of current through the capillary and prohibiting bands from reaching the detector. We also developed the concept of preinjection of samples, for which the sample plate is mock injected onto matrix from the previous run to reduce the number of small ions that out-compete DNA sequencing fragments for injection. Subsequently, the samples are injected a second time onto freshly injected matrix, electrophoresed, and detected as usual. In general, we favor ethanol precipitation of reaction products over other cleanup methods for its ease of performance in a 96-well format and because it is cheap, technically simple, and easily performed in a highthroughput production setting. Specifically, we precipitate with 100% ethanol in the presence of ammonium or sodium acetate and use two 70% washes to help eliminate excess salt ions that can interfere with electrokinetic injection and to reduce the intensity of free dye terminator artifacts. Column cleanup methods have been investigated but have proven too expensive and irreproducible in our mock production studies. However, for projects in which unincorporated dye terminator signals will interfere with analysis, column cleanup is preferred. We also routinely resuspend precipitated samples in deionized water (as opposed to formamide-based solutions) before loading, because this provides the maximum yield of resuspended fragments from the pellet. On the 3700, samples resuspended only in water typically evaporate during the preceding runs, and we add one half of the final volume as deionized formamide (after the initial water resuspension) to prevent complete evaporation. On the MegaBACE, the addition of one half of the final volume as 0.2% deionized hydroxyethyl cellulose helps to prevent most of the overinjection failures that occur with LPA, presumably by acting as a molecular weight sieve. Instrumentation Stability There are two phases of instrument stability to be considered. The first phase is stability for a newly released instrument, which is typically poor, because a variety of design, manufacturing, installation, and shipping issues are being sorted out. Both the MegaBACE and the 3700 have lived up to expectations with regard to this phase of stability, with each exhibiting a unique set of challenges. The second phase is long-term stability, which is assessed after the manufacturer has overcome most of the aforementioned issues. Although not all instruments successfully transition to the second phase of stability, the MegaBACE has entered that phase. We are evaluating these units with respect to their percentage of up time over the ensuing months. So far, the stability has been excellent, approaching more than 95% up time over the past 8 months. The 3700, because of its recent introduction, is in the initial phase of stability assessment. Capillary Lifetime A major cost-related issue in capillary sequencers is the expected lifetime of the capillary arrays. In the 3700, the capillaries are all part of a single bundle of 110 uncoated, fused silica capillaries, consisting of 96 primary capillaries, 8 reserve capillaries, and 6 sheath-flow guide capillaries. The reserve capillaries can be designated for use in the event that between one and eight of the primary capillaries is exhibiting substandard performance. In the MegaBACE, the capillaries are bundled together as sets of 16 coated, fused silica capillaries each, such that 6 separate array bundles comprise the 96. Whether an internal capillary coating is required is a function of the separation matrix used and directly affects the potential capillary lifetime. Intuitively, uncoated capillaries should last significantly longer than coated capillaries because the coating will wear away with repeated usage. As such, the specifications for Mega- BACE capillaries are a lifetime of up to 100 runs and for 3700 capillaries up to 300 runs. Perkin-Elmer reports that flushing capillaries with a nitric acid wash can regenerate them for up to 100 additional runs. How does specification compare with reality in our experience? Our records for MegaBACE capillary arrays indicate that, on average, 170 runs can be obtained from a set of 16 capillaries before the resolution, judged by analysis of a standard control reaction, falls below the manufacturer s specification of 550 bases. Although our experience with the 3700 is much less extensive, we have not yet achieved 300 uses from an array of capillaries before a similar assessment process indicates that resolution of sequencing reaction fragments falls below 300 bases. Nitric acid treatment of these capillaries has not successfully restored them to function in a limited number of attempts on different arrays, but recent 142 JOURNAL OF BIOMOLECULAR TECHNIQUES, VOLUME 10, ISSUE 3, SEPTEMBER 1999
7 CAPILLARY PLATFORMS FOR DNA SEQUENCING experience has taught us that several runs may be required before the acid-stripped capillaries again provide optimal performance. Matrix Performance The separation matrix is an integral part of the capillary sequencer and most of our development efforts have centered on overcoming the undesirable aspects of either matrix formulation. In the case of LPA, the matrix is extremely viscous and in our hands demonstrates the most potential for routinely obtaining sequences that span greater than 700 high-quality bases. Among its undesirable aspects, the pore size (among other physical characteristics) probably exacerbates the tendency of this matrix to overinject or clog with template molecules, proteins, or other species that out-compete the sequencing reaction fragments for loading. The incidence of clogging is undoubtedly a consequence of our choice to use lower purity template preparation methods, because our early experience with loading reactions obtained from polymerase chain reaction generated templates did not indicate a high percentage of failure due to clogs. The addition of hydroxyethyl cellulose before loading reactions on the MegaBACE significantly reduces or eliminates this problem. The POP6 matrix, having different physical characteristics from LPA, does not tend to clog, but it exhibits lower resolution potential. The modifications that we have made to run parameters, shown in Table 1, have primarily been aimed at improving POP6 resolution, although we have only achieved an average of 500 high-quality bases for the Big Dye terminator chemistry with these in place. throughput, infrastructure considerations may include a system to monitor capillary arrays for number of uses and performance, scheduled focusing assays, scheduled replacement of arrays on a rotating basis, and clearing of disk-archived data from hard drive storage. For the MegaBACE, a rotating supply of nitrogen tanks must be kept on hand. Some laboratories have reduced this need somewhat by using compressed air in place of a low-pressure nitrogen tank for driving the anode and cathode stages. For the 3700, routine nitric acid washes of capillaries may become necessary. In practice, these types of activities are best performed by a dedicated group that has the appropriate monitoring tools in place and is fully experienced in capillary replacement techniques and required postinstallation verification procedures. CONCLUSION This review was intended to be a comprehensive, unbiased report on our experience with two commercially available capillary sequencers, the MegaBACE 1000 and the Perkin-Elmer Potential buyers of capillary instrumentation should fully investigate the capabilities, specifications, and throughput of an instrument in light of the type of sequencing projects to be performed before a decision to purchase is made. ACKNOWLEDGMENTS The author wishes to thank John Bashkin and Jackie Snider for critical reading of the manuscript. Thanks also are extended to two valued collaborators, John Bashkin and Paul McEwan. Bob Waterston and Rick Wilson have provided encouragement and enlightenment, as always. INFRASTRUCTURE TO SUPPORT CAPILLARY INSTRUMENTS Labs that are planning to have multiple capillary platforms need to think carefully about the necessary infrastructure to support these instruments, because it is significantly different from that required for slab gel sequencers. Depending on the laboratory size and REFERENCES 1. Huang XC, Quesada MA, Mathies RA. Capillary array electrophoresis using laser-excited confocal fluorescence detection. Anal Chem 1992;64: Marsh M, Tu O, Dolnik V, et al. High-throughput DNA sequencing on a capillary array electrophoresis system. J Capillary Electrophor 1997;4: Kambara H, Takahashi S. Multiple-sheathflow capillary array DNA analyser. Nature 1993;361: JOURNAL OF BIOMOLECULAR TECHNIQUES, VOLUME 10, ISSUE 3, SEPTEMBER
CHAPTER 5 Troubleshooting DNA Sequencing Data
CHAPTER 5 Troubleshooting DNA Sequencing Data Instrument Artifacts Failed Injection High Background Color Balance Biased Reptation Electrophoresis Artifacts Weak Signal Overloading Current Fluctuations
More informationTroubleshooting Sequencing Data
Troubleshooting Sequencing Data Troubleshooting Sequencing Data No recognizable sequence (see page 7-10) Insufficient Quantitate the DNA. Increase the amount of DNA in the sequencing reactions. See page
More informationDNA Sequencing Setup and Troubleshooting
DNA Sequencing Setup and Troubleshooting Lara Cullen, PhD Scientific Applications Specialist Australia and New Zealand Reviewing Sequencing Data Review the Electropherogram Review the Raw Data (Signal
More informationSanger Sequencing and Quality Assurance. Zbigniew Rudzki Department of Pathology University of Melbourne
Sanger Sequencing and Quality Assurance Zbigniew Rudzki Department of Pathology University of Melbourne Sanger DNA sequencing The era of DNA sequencing essentially started with the publication of the enzymatic
More informationArtisan Scientific is You~ Source for: Quality New and Certified-Used/Pre:-awned ECJuiflment
Looking for more information? Visit us on the web at http://www.artisan-scientific.com for more information: Price Quotations Drivers Technical Specifications. Manuals and Documentation Artisan Scientific
More informationZR-96 DNA Sequencing Clean-up Kit Catalog Nos. D4052 & D4053
INSTRUCTION MANUAL ZR-96 DNA Sequencing Clean-up Kit Catalog Nos. D4052 & D4053 Highlights Simple 10 Minute Bind, Wash, Elute Procedure Flexible 15-20 µl Elution Volumes Allow for Direct Loading of Samples
More informationDNA Separation Methods. Chapter 12
DNA Separation Methods Chapter 12 DNA molecules After PCR reaction produces many copies of DNA molecules Need a way to separate the DNA molecules from similar sized molecules Only way to genotype samples
More informationAutomated DNA Sequencing. Chemistry Guide
Automated DNA Sequencing Chemistry Guide Copyright 2000, Applied Biosystems For Research Use Only. Not for use in diagnostic procedures. ABI PRISM and its design, Applied Biosystems, and MicroAmp are registered
More informationZR DNA Sequencing Clean-up Kit
INSTRUCTION MANUAL ZR DNA Sequencing Clean-up Kit Catalog Nos. D40 & D4051 Highlights Simple 2 Minute Bind, Wash, Elute Procedure Flexible 6-20 µl Elution Volumes Allow for Direct Loading of Samples with
More informationAurora Forensic Sample Clean-up Protocol
Aurora Forensic Sample Clean-up Protocol 106-0008-BA-D 2015 Boreal Genomics, Inc. All rights reserved. All trademarks are property of their owners. http://www.borealgenomics.com support@borealgenomics.com
More informationObjectives: Vocabulary:
Introduction to Agarose Gel Electrophoresis: A Precursor to Cornell Institute for Biology Teacher s lab Author: Jennifer Weiser and Laura Austen Date Created: 2010 Subject: Molecular Biology and Genetics
More informationDNA SEQUENCING (using an ABI automated sequencer)
DNA SEQUENCING (using an ABI automated sequencer) OBTECTIVE: To label and separate DNA fragments varying by single nucleotides, in order to determine the sequence of nucleotides. INTRODUCTION: Determination
More informationTroubleshooting Guide for DNA Electrophoresis
Troubleshooting Guide for Electrophoresis. ELECTROPHORESIS Protocols and Recommendations for Electrophoresis electrophoresis problem 1 Low intensity of all or some bands 2 Smeared bands 3 Atypical banding
More informationAutomotive Applications of 3D Laser Scanning Introduction
Automotive Applications of 3D Laser Scanning Kyle Johnston, Ph.D., Metron Systems, Inc. 34935 SE Douglas Street, Suite 110, Snoqualmie, WA 98065 425-396-5577, www.metronsys.com 2002 Metron Systems, Inc
More informationSanger Sequencing: Sample Preparation Guide
Sanger Sequencing: Sample Preparation Guide Use this as a guide to prepare your samples for Sanger sequencing at AGRF CONTENTS 1 Overview... 2 1.1 Capillary Separation (CS) or electrophoretic separation
More informationFOR REFERENCE PURPOSES
BIOO LIFE SCIENCE PRODUCTS FOR REFERENCE PURPOSES This manual is for Reference Purposes Only. DO NOT use this protocol to run your assays. Periodically, optimizations and revisions are made to the kit
More informationAutomated High Throughput Purification of BigDye TM Terminator Fluorescent DNA Sequencing Reactions Using Wizard MagneSil TM Paramagnetic Particles
Automated High Throughput Purification of BigDye TM Terminator Fluorescent DNA Sequencing Reactions Using Wizard MagneSil TM Paramagnetic Particles Paul Otto*, Brad Larson and Steve Krueger Abstract We
More informationElectrophoresis, cleaning up on spin-columns, labeling of PCR products and preparation extended products for sequencing
Electrophoresis, cleaning up on spin-columns, labeling of PCR products and preparation extended products for sequencing PAGE electrophoresis Polyacrylamide gel electrophoresis (PAGE) is used for separating
More informationWestern Blot Analysis with Cell Samples Grown in Channel-µ-Slides
Western Blot Analysis with Cell Samples Grown in Channel-µ-Slides Polyacrylamide gel electrophoresis (PAGE) and subsequent analyses are common tools in biochemistry and molecular biology. This Application
More informationFluorescent Array Imaging Reader
Fluorescent Array Imaging Reader Multiplex Enabled For 2-Color Fluorescent Microarrays Extended Dynamic Range Automated Spot Analysis Compact and Affordable Microarray Analysis With FLAIR FLAIR Sensovation
More informationTroubleshooting Overview
Overview This chapter provides information for troubleshooting automated DNA sequencing results from capillary electrophoresis runs. Assumptions Using Controls suggestions listed in this chapter assume
More informationLab 5: DNA Fingerprinting
Lab 5: DNA Fingerprinting You are about to perform a procedure known as DNA fingerprinting. The data obtained may allow you to determine if the samples of DNA that you will be provided with are from the
More informationA Brief Guide to Interpreting the DNA Sequencing Electropherogram Version 3.0
A Brief Guide to Interpreting the DNA Sequencing Electropherogram Version 3.0 Plant-Microbe Genomics Facility The Ohio State University 484 W.12 th Ave., Columbus, OH 43210 Ph: 614/247-6204 FAX: 614/247-8696
More informationReal-Time PCR Vs. Traditional PCR
Real-Time PCR Vs. Traditional PCR Description This tutorial will discuss the evolution of traditional PCR methods towards the use of Real-Time chemistry and instrumentation for accurate quantitation. Objectives
More informationApplication Guide... 2
Protocol for GenomePlex Whole Genome Amplification from Formalin-Fixed Parrafin-Embedded (FFPE) tissue Application Guide... 2 I. Description... 2 II. Product Components... 2 III. Materials to be Supplied
More informationAxyPrep TM Mag PCR Clean-up Protocol
AxyPrep TM Mag PCR Clean-up Protocol Intro The AxyPrep Mag PCR Clean-up kit utilizes a unique paramagnetic bead technology for rapid, high-throughput purification of PCR amplicons. Using this kit, PCR
More informationUltraClean PCR Clean-Up Kit
UltraClean PCR Clean-Up Kit Catalog No. Quantity 12500-50 50 Preps 12500-100 100 Preps 12500-250 250 Preps Instruction Manual Please recycle Version: 02212013 1 Table of Contents Introduction... 3 Protocol
More informationUltraClean Soil DNA Isolation Kit
PAGE 1 UltraClean Soil DNA Isolation Kit Catalog # 12800-50 50 preps New improved PCR inhibitor removal solution (IRS) included Instruction Manual (New Alternative Protocol maximizes yields) Introduction
More information7 Electrophoresis. µ proportional to Q
7 Electrophoresis Objectives: A) To perform agarose gel electrophoresis of the proteins isolated in last week's experiment and B) to interpret the banding patterns produced by these proteins. Introduction:
More informationAGAROSE GEL ELECTROPHORESIS:
AGAROSE GEL ELECTROPHORESIS: BEST PRACTICES (BACK TO THE BASICS) Unit of Tropical Laboratory Medicine April 2009 Marcella Mori WORKFLOW OF AGAROSE GEL ELECTROPHORESIS: THREE STEPS Agarose gel electrophoresis
More informationProcedures For DNA Sequencing
Procedures For DNA Sequencing Plant-Microbe Genomics Facility (PMGF) Ohio State University 420 Biological Sciences Building 484 W. 12th Ave., Columbus OH 43210 Telephone: 614/247-6204 FAX: 614/292-6337
More informationAMINO ACID ANALYSIS By High Performance Capillary Electrophoresis
AMINO ACID ANALYSIS By High Performance Capillary Electrophoresis Analysis of Amino Acid Standards Label free analysis using the HPCE-512 ABSTRACT Capillary electrophoresis using indirect UV detection
More informationDNA Detection. Chapter 13
DNA Detection Chapter 13 Detecting DNA molecules Once you have your DNA separated by size Now you need to be able to visualize the DNA on the gel somehow Original techniques: Radioactive label, silver
More informationIntroduction. Preparation of Template DNA
Procedures and Recommendations for DNA Sequencing at the Plant-Microbe Genomics Facility Ohio State University Biological Sciences Building Room 420, 484 W. 12th Ave., Columbus OH 43210 Telephone: 614/247-6204;
More informationInc. Wuhan. Quantity Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4 Standard (liquid) 2
Uscn Life Science Inc. Wuhan Website: www.uscnk.com Phone: +86 27 84259552 Fax: +86 27 84259551 E-mail: uscnk@uscnk.com ELISA Kit for Human Prostaglandin E1(PG-E1) Instruction manual Cat. No.: E0904Hu
More informationExperiment #5: Qualitative Absorption Spectroscopy
Experiment #5: Qualitative Absorption Spectroscopy One of the most important areas in the field of analytical chemistry is that of spectroscopy. In general terms, spectroscopy deals with the interactions
More informationCanine creatine kinase MB isoenzyme (CK-MB)ELISA Kit
Canine creatine kinase MB isoenzyme (CK-MB)ELISA Kit Catalog No. CSB-E15852c (96T) This immunoassay kit allows for the in vitro quantitative determination of canine CK-MB concentrations in serum and plasma.
More informationDye-Blob message: Example: Generally, this is due to incomplete excess dye removal of the cycle sequence reaction.
When sequence data is uploaded to ilab, an email is sent notifying the user that data is ready. The staff of the DNA facility has the ability to edit this message to include specific remarks about how
More informationPreciseTM Whitepaper
Precise TM Whitepaper Introduction LIMITATIONS OF EXISTING RNA-SEQ METHODS Correctly designed gene expression studies require large numbers of samples, accurate results and low analysis costs. Analysis
More informationHuman Free Testosterone(F-TESTO) ELISA Kit
Human Free Testosterone(F-TESTO) ELISA Kit Catalog Number. MBS700040 For the quantitative determination of human free testosterone(f-testo) concentrations in serum, plasma. This package insert must be
More informationRealStar HBV PCR Kit 1.0 11/2012
RealStar HBV PCR Kit 1.0 11/2012 RealStar HBV PCR Kit 1.0 For research use only! (RUO) Product No.: 201003 96 rxns INS-201000-GB-02 Store at -25 C... -15 C November 2012 altona Diagnostics GmbH Mörkenstraße
More informationDNA Core Facility: DNA Sequencing Guide
DNA Core Facility: DNA Sequencing Guide University of Missouri-Columbia 216 Life Sciences Center Columbia, MO 65211 http://biotech.missouri.edu/dnacore/ Table of Contents 1. Evaluating Sequencing Data..
More informationBigDye Terminator v3.1 Cycle Sequencing Kit. Protocol. DRAFT August 27, 2002 12:32 pm, 4337035A_v3.1Title.fm
BigDye Terminator v3.1 Cycle Sequencing Kit Protocol August 27, 2002 12:32 pm, 4337035A_v3.1Title.fm Copyright 2002, Applied Biosystems. All rights reserved. For Research Use Only. Not for use in diagnostic
More informationElemental Analyses by ICP-AES
Elemental Analyses by ICP-AES Henry Gong, Senior Analytical Chemist September 10, 2008 ICP-AES inductively coupled plasma atomic emission spectrophotometry Electrons of an atom absorb energy and jump to
More informationVLLM0421c Medical Microbiology I, practical sessions. Protocol to topic J10
Topic J10+11: Molecular-biological methods + Clinical virology I (hepatitis A, B & C, HIV) To study: PCR, ELISA, your own notes from serology reactions Task J10/1: DNA isolation of the etiological agent
More informationApplied Biosystems 3500/3500xL Genetic Analyzer
QUICK REFERENCE Applied Biosystems 3500/3500xL Genetic Analyzer with 3500 Series Data Collection Software 3 Catalog Number 4405186, 4405187 Pub. No. 100026299 Rev. A Note: For safety and biohazard guidelines,
More informationEZ-PAGE Electrophoresis System USER MANUAL
EZ-PAGE Electrophoresis System USER MANUAL Table of Contents Safety Information.. 2 Product Description... 2 Product Contents..... 3 Specifications & Storage Conditions.. 3 Product Use..... 3 Getting Started
More informationClassic Immunoprecipitation
292PR 01 G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name Classic Immunoprecipitation Utilizes Protein A/G Agarose for Antibody Binding (Cat.
More informationab185915 Protein Sumoylation Assay Ultra Kit
ab185915 Protein Sumoylation Assay Ultra Kit Instructions for Use For the measuring in vivo protein sumoylation in various samples This product is for research use only and is not intended for diagnostic
More informationHow is genome sequencing done?
How is genome sequencing done? Using 454 Sequencing on the Genome Sequencer FLX System, DNA from a genome is converted into sequence data through four primary steps: Step One DNA sample preparation; Step
More informationDevelop a Quantitative Analytical Method for low (» 1 ppm) levels of Sulfate
Cantaurus, Vol. 7, 5-8, May 1999 McPherson College Division of Science and Technology Develop a Quantitative Analytical Method for low (» 1 ppm) levels of Sulfate Janet Bowen ABSTRACT Sulfate is used in
More informationUltraClean Forensic DNA Isolation Kit (Single Prep Format)
UltraClean Forensic DNA Isolation Kit (Single Prep Format) Catalog No. Quantity 14000-10 10 preps 14000-S 1 prep Instruction Manual Please recycle Version: 10302012 1 Table of Contents Introduction...
More informationINSERT COMPANY LOGO HERE
2013 2014 INSERT COMPANY LOGO HERE 2014 2013 North American Sample SSL Certificate Analysis New Product Product Leadership Innovation Award Award New Product Innovation Award Sample Analysis North America,
More informationEQUIPMENT CALIBRATION/MAINTENANCE
EQUIPMENT CALIBRATION/MAINTENANCE The following chart provides a guide for calibration of standard equipment used in the Fish Health Laboratory. All Intervals suggested are subject to individual manufacturer
More informationEnvironmental Water Testing: Surface Water, Groundwater, Hard Water, Wastewater, & Seawater
Document: AND Sol Env 08 2013 Environmental Water Testing: Surface Water, Groundwater, Hard Water, Wastewater, & Seawater Matrix specific sample preparation and testing methods for environmental waters
More informationUniversidade Estadual de Maringá
Universidade Estadual de Maringá Disciplina: Biologia Molecular Sequenciamento de ácidos nucléicos Profa. Dra. Maria Aparecida Fernandez Maxan e Gilbert - quebra química Berg, Gilbert and Sanger dideoxinucleotideos
More informationThe Biotechnology Education Company
EDVTEK P.. Box 1232 West Bethesda, MD 20827-1232 The Biotechnology 106 EDV-Kit # Principles of DNA Sequencing Experiment bjective: The objective of this experiment is to develop an understanding of DNA
More informationHidex Sense. Hidex Triathler Multi-technology Microplate Reader. Hidex AMG. Consumables. Hidex 300 SL. www.lablogic.com.
Service and Support Users of our systems can benefit from our comprehensive, fully inclusive service and support. We provide complete service and support for all of our customers to give reassurance that
More informationSequencing Guidelines Adapted from ABI BigDye Terminator v3.1 Cycle Sequencing Kit and Roswell Park Cancer Institute Core Laboratory website
Biomolecular Core Facility AI Dupont Hospital for Children, Rockland Center One, Room 214 Core: (302) 651-6712, Office: (302) 651-6707, mbcore@nemours.org Katia Sol-Church, Ph.D., Director Jennifer Frenck
More informationINSTRUCTIONS 56-1190-98. Edition AC
Sephacryl S-100 High Resolution Sephacryl S-200 High Resolution Sephacryl S-300 High Resolution Sephacryl S-400 High Resolution Sephacryl S-500 High Resolution INSTRUCTIONS Sephacryl High Resolution chromatography
More informationDNA Sequence Analysis
DNA Sequence Analysis Two general kinds of analysis Screen for one of a set of known sequences Determine the sequence even if it is novel Screening for a known sequence usually involves an oligonucleotide
More informationTechnical Note. Roche Applied Science. No. LC 19/2004. Color Compensation
Roche Applied Science Technical Note No. LC 19/2004 Purpose of this Note Color The LightCycler System is able to simultaneously detect and analyze more than one color in each capillary. Due to overlap
More informationGeospiza s Finch-Server: A Complete Data Management System for DNA Sequencing
KOO10 5/31/04 12:17 PM Page 131 10 Geospiza s Finch-Server: A Complete Data Management System for DNA Sequencing Sandra Porter, Joe Slagel, and Todd Smith Geospiza, Inc., Seattle, WA Introduction The increased
More informationMultiQuant Software 2.0 for Targeted Protein / Peptide Quantification
MultiQuant Software 2.0 for Targeted Protein / Peptide Quantification Gold Standard for Quantitative Data Processing Because of the sensitivity, selectivity, speed and throughput at which MRM assays can
More informationORGANIC SAMPLE PREPARATION
ORGANIC SAMPLE PREPARATION W W W.LA BT E C H S R L.CO M WSPE MANUAL VACUUM MANIFOLD SPE Process control of the flow rate is critical to guarantee reproducible extractions. Differently then any other systems,
More informationExpressArt Bacterial H-TR cdna synthesis kit. With extreme selectivity against rrnas
ExpressArt Bacterial H-TR cdna synthesis kit With extreme selectivity against rrnas suitable for 2 to 4 µg total RNA Catalogue No. 8004-A30 (30 rxns) Reagents Materials are provided for 30 cdna synthesis
More informationLecture 20: Scanning Confocal Microscopy (SCM) Rationale for SCM. Principles and major components of SCM. Advantages and major applications of SCM.
Lecture 20: Scanning Confocal Microscopy (SCM) Rationale for SCM. Principles and major components of SCM. Advantages and major applications of SCM. Some limitations (disadvantages) of NSOM A trade-off
More informationClinical trial for 510(k) clearance
CAPILLARY ELECTROPHORESIS Clinical trial for 510(k) clearance The 3500 and 3500xL Dx Genetic Analyzers CS2 CO19459 Molokai- Instrument prelaunch brochure.indd 1 The next step in molecular medicine 3500
More informationBiotracker TM A Laboratory Information Management System By Ocimum Biosolutions
Biotracker TM A Laboratory Information Management System By Ocimum Biosolutions 1 TABLE OF CONTENTS 1.0 EXECUTIVE SUMMARY... 2 2.0 INTRODUCTION... 2 3.0 BIOTRACKER TM GENERAL FEATURES... 4 3.1 LABORATORY
More informationIntroduction to flow cytometry
Introduction to flow cytometry Flow cytometry is a popular laser-based technology. Discover more with our introduction to flow cytometry. Flow cytometry is now a widely used method for analyzing the expression
More informationWelcome to Pacific Biosciences' Introduction to SMRTbell Template Preparation.
Introduction to SMRTbell Template Preparation 100 338 500 01 1. SMRTbell Template Preparation 1.1 Introduction to SMRTbell Template Preparation Welcome to Pacific Biosciences' Introduction to SMRTbell
More informationISOLATE II PCR and Gel Kit. Product Manual
ISOLATE II PCR and Gel Kit Product Manual 2 Product Manual www.bioline.com/isolate PCR and Gel Kit ISOLATE II PCR and Gel Kit ISOLATE II PCR and Gel Kit 1 Kit contents 04 2 Description 04 3 Storage 04
More informationThe fastest spin-column based procedure for purifying up to 10 mg of ultra-pure endotoxin-free transfection-grade plasmid DNA.
INSTRUCTION MANUAL ZymoPURE Plasmid Gigaprep Kit Catalog Nos. D4204 (Patent Pending) Highlights The fastest spin-column based procedure for purifying up to 10 mg of ultra-pure endotoxin-free transfection-grade
More informationNATIONAL GENETICS REFERENCE LABORATORY (Manchester)
NATIONAL GENETICS REFERENCE LABORATORY (Manchester) MLPA analysis spreadsheets User Guide (updated October 2006) INTRODUCTION These spreadsheets are designed to assist with MLPA analysis using the kits
More informationDNA Sequencing Overview
DNA Sequencing Overview DNA sequencing involves the determination of the sequence of nucleotides in a sample of DNA. It is presently conducted using a modified PCR reaction where both normal and labeled
More informationRECITATION NOTES FOR EXPERIMENT # 5 A&B THIN LAYER CHROMATOGRAPHY
RECITATION NOTES FOR EXPERIMENT # 5 A&B THIN LAYER CHROMATOGRAPHY Have your lab textbook available for quick reference to specific pages, indicated in red. BASIC PRINCIPLES OF CHROMATOGRAPHY Chromatography
More informationAn In-Gel Digestion Protocol
An In-Gel Digestion Protocol This protocol describes the digestion of a protein present in an SDS-PAGE gel band with trypsin. The band can be taken from either a 1D or 2D electrophoresis gel. Reagents
More informationElectrochemistry Revised 04/29/15
INTRODUCTION TO ELECTROCHEMISTRY: CURRENT, VOLTAGE, BATTERIES, & THE NERNST EQUATION Experiment partially adapted from J. Chem. Educ., 2008, 85 (8), p 1116 Introduction Electrochemical cell In this experiment,
More informationAB SCIEX TOF/TOF 4800 PLUS SYSTEM. Cost effective flexibility for your core needs
AB SCIEX TOF/TOF 4800 PLUS SYSTEM Cost effective flexibility for your core needs AB SCIEX TOF/TOF 4800 PLUS SYSTEM It s just what you expect from the industry leader. The AB SCIEX 4800 Plus MALDI TOF/TOF
More informationApplication Note. Single Cell PCR Preparation
Application Note Single Cell PCR Preparation From Automated Screening to the Molecular Analysis of Single Cells The AmpliGrid system is a highly sensitive tool for the analysis of single cells. In combination
More informationChapter 3 Contd. Western blotting & SDS PAGE
Chapter 3 Contd. Western blotting & SDS PAGE Western Blot Western blots allow investigators to determine the molecular weight of a protein and to measure relative amounts of the protein present in different
More informationMeso Scale Discovery. WINDOWS is a registered trademark of Microsoft Corporation
Meso Scale Discovery M S D M S D Meso Scale Discovery, a division of Meso Scale Diagnostics, LLC. (MSD) 9238 Gaither Rd, Gaithersburg, MD 20877 Phone: 240.631.2522 Fax: 240.632.2219 email: sales@meso-scale.com
More informationElectrospray Ion Trap Mass Spectrometry. Introduction
Electrospray Ion Source Electrospray Ion Trap Mass Spectrometry Introduction The key to using MS for solutions is the ability to transfer your analytes into the vacuum of the mass spectrometer as ionic
More informationab185916 Hi-Fi cdna Synthesis Kit
ab185916 Hi-Fi cdna Synthesis Kit Instructions for Use For cdna synthesis from various RNA samples This product is for research use only and is not intended for diagnostic use. Version 1 Last Updated 1
More informationWaters Corporation. Waters 2690/5 USER & TROUBLESHOOTING GUIDE
Waters Corporation Waters 2690/5 USER & TROUBLESHOOTING GUIDE Contents 2690/5 Theory Setup procedures. Troubleshooting the 2690/5 User maintenance of the 2690/5 Spare Parts 2 2690/5 Theory 2690/5 Solvent
More informationABI PRISM 3100 Genetic Analyzer. User s Manual
ABI PRISM 3100 Genetic Analyzer User s Manual Copyright 2001, Applied Biosystems For Research Use Only. Not for use in diagnostic procedures. NOTICE TO PURCHASER This instrument, Serial No., is Authorized
More informationPowerFecal DNA Isolation Kit
PowerFecal DNA Isolation Kit Catalog No. Quantity 12830-50 50 Preps Instruction Manual Inhibitor Removal Technology (IRT) is a registered trademark of MO BIO Laboratories, Inc. and is covered by the following
More informationUniversity of Wisconsin Chemistry 524 Spectroscopic Applications (GFAA, ICP, UV/Vis, Fluorescence)
University of Wisconsin Chemistry 524 Spectroscopic Applications (GFAA, ICP, UV/Vis, Fluorescence) For this laboratory exercise, you will explore a variety of spectroscopic methods used in an analytical
More informationSDS-PAGE Protocol Mutated from the SDS-PAGE protocol written by the Lord of the Flies
SDS-PAGE Protocol Mutated from the SDS-PAGE protocol written by the Lord of the Flies Pouring the resolving gel 1. Clean glass plates with soap and water, then with ethanol. Assemble the glass plates and
More informationRESTRICTION ENZYME ANALYSIS OF DNA
University of Massachusetts Medical School Regional Science Resource Center SUPPORTING MATHEMATICS, SCIENCE AND TECHNOLOGY EDUCATION 222 Maple Avenue, Stoddard Building Shrewsbury, MA 01545-2732 508.856.5097
More informationArtisanLink Staining System is an automated special stains slide
Technical Tips Tips on using the ArtisanLink Special Staining System Jamie Nowacek, BS, HT(ASCP) CM, QIHC, PMP Dako North America, Inc. Carpinteria, CA, USA ArtisanLink Staining System is an automated
More informationMouse Keyhole Limpet Hemocyanin antibody(igm) ELISA Kit
Mouse Keyhole Limpet Hemocyanin antibody(igm) ELISA Kit Catalog No. MBS702810 (96 tests) This immunoassay kit allows for the in vitro semi-quantitative determination of mouse KLH(IgM)antibody concentrations
More informationCrime Scenes and Genes
Glossary Agarose Biotechnology Cell Chromosome DNA (deoxyribonucleic acid) Electrophoresis Gene Micro-pipette Mutation Nucleotide Nucleus PCR (Polymerase chain reaction) Primer STR (short tandem repeats)
More informationHiPer RT-PCR Teaching Kit
HiPer RT-PCR Teaching Kit Product Code: HTBM024 Number of experiments that can be performed: 5 Duration of Experiment: Protocol: 4 hours Agarose Gel Electrophoresis: 45 minutes Storage Instructions: The
More informationTIANquick Mini Purification Kit
TIANquick Mini Purification Kit For purification of PCR products, 100 bp to 20 kb www.tiangen.com TIANquick Mini Purification Kit (Spin column) Cat no. DP203 Kit Contents Contents Buffer BL Buffer PB Buffer
More informationPicoMaxx High Fidelity PCR System
PicoMaxx High Fidelity PCR System Instruction Manual Catalog #600420 (100 U), #600422 (500 U), and #600424 (1000 U) Revision C Research Use Only. Not for Use in Diagnostic Procedures. 600420-12 LIMITED
More informationChemistry 321, Experiment 8: Quantitation of caffeine from a beverage using gas chromatography
Chemistry 321, Experiment 8: Quantitation of caffeine from a beverage using gas chromatography INTRODUCTION The analysis of soft drinks for caffeine was able to be performed using UV-Vis. The complex sample
More informationAutomated Library Preparation for Next-Generation Sequencing
Buyer s Guide: Automated Library Preparation for Next-Generation Sequencing What to consider as you evaluate options for automating library preparation. Yes, success can be automated. Next-generation sequencing
More informationCONFIRMATION OF ZOLPIDEM BY LIQUID CHROMATOGRAPHY MASS SPECTROMETRY
CONFIRMATION OF ZOLPIDEM BY LIQUID CHROMATOGRAPHY MASS SPECTROMETRY 9.1 POLICY This test method may be used to confirm the presence of zolpidem (ZOL), with diazepam-d 5 (DZP-d 5 ) internal standard, in
More informationRNA Extraction and Quantification, Reverse Transcription, and Real-time PCR (q-pcr)
RNA Extraction and Quantification, Reverse Transcription, and Real-time Preparation of Samples Cells: o Remove media and wash cells 2X with cold PBS. (2 ml for 6 well plate or 3 ml for 6cm plate) Keep
More information