DNA mini preparation
|
|
- Alexander McDonald
- 7 years ago
- Views:
Transcription
1 1 DNA mini preparation Mini-Preparation of Plasmid DNA Mini-Prep = The isolation and purification of plasmid DNA from bacteria. In order to use a vector for cloning, sequencing, etc., it is necessary to isolate the vector in a highly purified form. Many companies now sell kits which provide all the solutions necessary for preparing DNA. Most Mini-Preparation techniques use the alkaline lysis protocol. Mini-prep
2 2 Mini-Preparation of Plasmid DNA Centrifugation - The process of separating substances by different densities by using a centrifuge. Supernatant Substances which are more dense settle to the bottom of the tube. Pellet the clump of cells and cellular debris found at the bottom of a centrifuge tube after centrifugation. Pellet Supernatant The nonpelleted solution left after centrifugation. Transfer 1.5 ml of the culture to a microfuge tube and pellet the cells for 1 minute at full speed (14,000 rpm) in the microcentrifuge.
3 3 Remove the growth medium by pouring out the supernatant. Gently tap the microcentrifuge tube on a paper towel to remove any residual supernatent. Resuspend the bacterial pellet in 200µl of solution I by pipetting up and down as well as vortexing. Solution I (resuspension buffer) Solution I contains three components: glucose, Tris and EDTA. Glucose and Tris are used to buffer the ph of the cell suspension. EDTA chelates divalent cations (ions with charges of +2) such as Mg ++. This helps break down the cell membrane and inactivate DNAase (enzymes which break down DNA)
4 4 Add 200 µl of Solution II, invert gently 10 times. Do not vortex!! This will shear the DNA and contaminate your DNA preps. Solution 2 (alakaline lysis buffer): P2 is the solution that will lyse the cells. The buffer contains.2m NaOH and 1% SDS. SDS is a detergent that will destroy the bacterial membrane. NaOH will denature the chromosomal DNA and the plasmid DNA. Add 400 µl of Solution III. Invert gently 10 times. Do not vortex. Solution 3 (neutralization buffer): Solution 3 consists of 5M KOAc, ph 4.8 KOAc will readjust the ph and allow only the plasmid DNA to reanneal. Because the chromosomal DNA is so large, it cannot reanneal properly under these conditions.
5 5 Centrifuge for 5 minutes at full speed in the microcentrifuge. A white pellet will form on the bottom and side of the tube after centrifugation. The plasmid DNA must now be purified away from the rest of the cytoplasm and concentrated. Affinity chromatography DNA selectively binds to the column. Pour the supernatant into the appropriately labeled spin column which has been inserted into a 2 ml collection tube.
6 6 Centrifuge for 1 minute at full speed, and drain the flow-through from the collection tube. Add 400 µl of Wash Buffer to the spin column contained in the 2 ml Collection Tube, centrifuge at full speed for 1 minute, and drain the flow through. Wash buffer Once the plasmid DNA has been added to the column, the wash buffer is used to wash away any unwanted material.
7 7 Place the spin column in a fresh 1.7 ml microcentrifuge tube and centrifuge again for 1 minute at full speed to remove any residual wash solution that might still be in the column. Place the spin column into an appropriately labeled 1.7 ml microcentrifuge tube and add 60 µl of EB (Elution buffer) to the column. Elute - Remove plasmid DNA from the column Centrifuge at full speed for 1 minute. Elution buffer EB removes the plasmid DNA from the column. It is important to keep this volume relatively small in order to have a reasonable DNA concentration. Autoclaved H 2 O can also be used to elute the DNA.
8 8 Remove the spin column from the labeled 1.7 ml microcentrifuge tube and close the lid on the tube tightly. Store it in a freezer box with your school number and your initials. Place the freezer box in the freezer. Enter the name and location of the plasmid in the clone log Book keeping!! Enter the clone name and prep date into Google Docs
LAB 11 PLASMID DNA MINIPREP
LAB 11 PLASMID DNA MINIPREP STUDENT GUIDE GOAL The objective of this lab is to perform extraction of plasmid DNA and analyze the results. OBJECTIVES After completion, the student should be able to: 1.
More informationHighPure Maxi Plasmid Kit
HighPure Maxi Plasmid Kit For purification of high pure plasmid DNA with high yields www.tiangen.com PP120109 HighPure Maxi Plasmid Kit Kit Contents Storage Cat.no. DP116 Contents RNaseA (100 mg/ml) Buffer
More informationLab 10: Bacterial Transformation, part 2, DNA plasmid preps, Determining DNA Concentration and Purity
Lab 10: Bacterial Transformation, part 2, DNA plasmid preps, Determining DNA Concentration and Purity Today you analyze the results of your bacterial transformation from last week and determine the efficiency
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 informationPlant Genomic DNA Extraction using CTAB
Plant Genomic DNA Extraction using CTAB Introduction The search for a more efficient means of extracting DNA of both higher quality and yield has lead to the development of a variety of protocols, however
More informationPurification of Plasmid DNA
Purification of Plasmid DNA Introduction: The growth of colonies on antibiotic medium provides phenotypic evidence that cells have been transformed. To confirm this at the genotypic level, plasmid DNA
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 information50 g 650 L. *Average yields will vary depending upon a number of factors including type of phage, growth conditions used and developmental stage.
3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com Phage DNA Isolation Kit Product # 46800, 46850 Product Insert
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 informationGRS Plasmid Purification Kit Transfection Grade GK73.0002 (2 MaxiPreps)
1 GRS Plasmid Purification Kit Transfection Grade GK73.0002 (2 MaxiPreps) (FOR RESEARCH ONLY) Sample : Expected Yield : Endotoxin: Format : Operation Time : Elution Volume : 50-400 ml of cultured bacterial
More informationGenolution Pharmaceuticals, Inc. Life Science and Molecular Diagnostic Products
Genolution Pharmaceuticals, Inc. Revolution through genes, And Solution through genes. Life Science and Molecular Diagnostic Products www.genolution1.com TEL; 02-3010-8670, 8672 Geno-Serum Hepatitis B
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 informationPCR and Sequencing Reaction Clean-Up Kit (Magnetic Bead System) 50 preps Product #60200
3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com PCR and Sequencing Reaction Clean-Up Kit (Magnetic Bead System)
More informationTransformAid Bacterial Transformation Kit
Home Contacts Order Catalog Support Search Alphabetical Index Numerical Index Restriction Endonucleases Modifying Enzymes PCR Kits Markers Nucleic Acids Nucleotides & Oligonucleotides Media Transfection
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 informationHow To Use An Enzymatics Spark Dna Sample Prep Kit For Ion Torrent
SPARK DNA Sample Prep Kit Ion Torrent (SPK0002-V08) Frequently Asked Questions Under what circumstances would I use SPARK DNA Sample Prep Kit for Ion Torrent? Enzymatics SPARK DNA Sample Prep Kit for Ion
More informationCLONING IN ESCHERICHIA COLI
CLONING IN ESCHERICHIA COLI Introduction: In this laboratory, you will carry out a simple cloning experiment in E. coli. Specifically, you will first create a recombinant DNA molecule by carrying out a
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 informationAutomation in Genomics High-throughput purification of nucleic acids from biological samples. Valentina Gualdi Operational Scientist PGP
Automation in Genomics High-throughput purification of nucleic acids from biological samples Valentina Gualdi Operational Scientist PGP OVERVIEW Nucleic acid purification technologies general aspects Genomic
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 informationDNA Isolation Kit for Cells and Tissues
DNA Isolation Kit for Cells and Tissues for 10 isolations of 00 mg each for tissue or 5 x 10 7 cultured cells Cat. No. 11 81 770 001 Principle Starting material Application Time required Results Benefits
More informationTransformation Protocol
To make Glycerol Stocks of Plasmids ** To be done in the hood and use RNase/DNase free tips** 1. In a 10 ml sterile tube add 3 ml autoclaved LB broth and 1.5 ul antibiotic (@ 100 ug/ul) or 3 ul antibiotic
More informationTECHNICAL BULLETIN. HIS-Select Nickel Affinity Gel. Catalog Number P6611 Storage Temperature 2 8 C
HIS-Select Nickel Affinity Gel Catalog Number P6611 Storage Temperature 2 8 C TECHNICAL BULLETIN Product Description HIS-Select Nickel Affinity Gel is an immobilized metalion affinity chromatography (IMAC)
More informationGenomic DNA Extraction Kit INSTRUCTION MANUAL
Genomic DNA Extraction Kit INSTRUCTION MANUAL Table of Contents Introduction 3 Kit Components 3 Storage Conditions 4 Recommended Equipment and Reagents 4 Introduction to the Protocol 4 General Overview
More informationIsolation and Purification of Total Genomic DNA from Gram-Negative Bacteria
Isolation and Purification of Total Genomic DNA from Gram-Negative Bacteria INTRODUCTION The isolation and purification of DNA from cells is one of the most common procedures in contemporary molecular
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 informationncounter Gene Expression Assay Manual Total RNA and Cell Lysate Protocols
ncounter Gene Expression Assay Manual Total RNA and Cell Lysate Protocols v.20090807 For research use only. Not for use in diagnostic procedures. Limited License Subject to the terms and conditions of
More informationInvestigating a Eukaryotic Genome: Cloning and Sequencing a Fragment of Yeast DNA
Investigating a Eukaryotic Genome: Cloning and Sequencing a Fragment of Yeast DNA Credits: This lab was created by Sarah C.R. Elgin and developed and written by Kathleen Weston-Hafer. Specific protocols
More informationELUTION OF DNA FROM AGAROSE GELS
ELUTION OF DNA FROM AGAROSE GELS OBTECTIVE: To isolate specific bands or regions of agarose-separated DNA for use in subsequent experiments and/or procedures. INTRODUCTION: It is sometimes necessary to
More informationHiPer Total RNA Extraction Teaching Kit
HiPer Total RNA Extraction Teaching Kit Product Code: HTBM012 Number of experiments that can be performed: 10 Duration of Experiment Protocol: 1 hour Agarose Gel Electrophoresis: 1 hour Storage Instructions:
More informationRNA PowerSoil Total RNA Isolation Kit Sample (Catalog No. 12866-S) Information for Ordering Product Catalog No. Quantity 12866-25 25 Preps
RNA PowerSoil Total RNA Isolation Kit Sample (Catalog No. 12866-S) Information for Ordering Product Catalog No. Quantity 12866-25 25 Preps Instruction Manual New protocol instructions: (Steps 3-5) Phenol:chloroform:isoamyl
More informationCHEF Genomic DNA Plug Kits Instruction Manual
CHEF Genomic DNA Plug Kits Instruction Manual Catalog Numbers 170-3591 170-3592 170-3593 For Technical Service Call Your Local Bio-Rad Office or in the U.S. Call 1-800-4BIORAD (1-800-424-6723) Table of
More informationMICB ABI PRISM 310 SEQUENCING GUIDE SEQUENCING OF PLASMID DNA
Plasmid DNA Preparation MICB ABI PRISM 310 SEQUENCING GUIDE SEQUENCING OF PLASMID DNA Introduction: I have always used the classic Alkaline Lysis miniprep method to isolate plasmid DNA. (See below) If
More informationHiPer Ion Exchange Chromatography Teaching Kit
HiPer Ion Exchange Chromatography Teaching Kit Product Code: HTC001 Number of experiments that can be performed: 5 Duration of Experiment: Protocol: 5-6 hours Storage Instructions: The kit is stable for
More informationFrozen-EZ Yeast Transformation II Catalog No. T2001
INSTRUCTIONS Frozen-EZ Yeast Transformation II Catalog No. T2001 Highlights High transformation efficiency that yields approximately 10 5-10 6 transformants per μg plasmid DNA (circular). Frozen storage
More informationAgencourt RNAdvance Blood Kit for Free Circulating DNA and mirna/rna Isolation from 200-300μL of Plasma and Serum
SUPPLEMENTAL PROTOCOL WHITE PAPER Agencourt RNAdvance Blood Kit for Free Circulating DNA and mirna/rna Isolation from 200-300μL of Plasma and Serum Bee Na Lee, Ph.D., Beckman Coulter Life Sciences Process
More informationRecommended Procedures for the Extraction of RNA. Jan Pedersen USDA, APHIS, VS, National Veterinary Services Laboratories, Ames, IA 50010
Recommended Procedures for the Extraction of RNA Jan Pedersen USDA, APHIS, VS, National Veterinary Services Laboratories, Ames, IA 50010 RNA Extraction Isolates RNA from other cellular components in the
More informationMagExtractor -Genome-
Instruction manual MagExtractor-Genome-0810 F0981K MagExtractor -Genome- NPK-101 100 preparations Store at 4 C Contents [1] Introduction [2] Components [3] Materials required [4] Protocol 1. Purification
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 informationUser s Manual. Bacteria Genomic DNA Kit. ExiPrepTM Bacteria Genomic DNA Kit K-3245. Version No.: 3.0 (2010-12-24) www.bioneer.com
ExiPrepTM Bacteria Genomic DNA Kit ExiPrepTM Bacteria Genomic DNA Kit Kit for the samples extraction using of bacteria ExiPrepTM genomic instrument DNA in various K-3245 Version No.: 3.0 (2010-12-24) User
More informationQIAGEN Supplementary Protocol
QIAGEN Supplementary Protocol Isolation of Peripheral Blood Mononuclear Cells (PBMC) and Purification of Total RNA from PBMC Using the RNeasy Micro or Mini Kit This protocol describes how to isolate PBMC
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 informationMolecular Biology Techniques: A Classroom Laboratory Manual THIRD EDITION
Molecular Biology Techniques: A Classroom Laboratory Manual THIRD EDITION Susan Carson Heather B. Miller D.Scott Witherow ELSEVIER AMSTERDAM BOSTON HEIDELBERG LONDON NEW YORK OXFORD PARIS SAN DIEGO SAN
More informationSV Total RNA Isolation System
TECHNICAL MANUAL SV Total RNA Isolation System Instruc ons for use of Products Z3105 and Z3100 Revised 8/13 TM048 SV Total RNA Isolation System All technical literature is available on the Internet at:
More informationDNA SPOOLING 1 ISOLATION OF DNA FROM ONION
DNA SPOOLING 1 ISOLATION OF DNA FROM ONION INTRODUCTION This laboratory protocol will demonstrate several basic steps required for isolation of chromosomal DNA from cells. To extract the chromosomal DNA,
More informationWizard DNA Clean-Up System INSTRUCTIONS FOR USE OF PRODUCT A7280.
Technical Bulletin Wizard DNA Clean-Up System INSTRUCTIONS FOR USE OF PRODUCT A7280. PRINTED IN USA. Revised 4/06 AF9TB141 0406TB141 Wizard DNA Clean-Up System All technical literature is available on
More informationImportant Note. September 2011
September 2011 Important Note Dear PAXgene Tissue mirna Kit user, When working with PAXgene Tissue treated samples of fibrous tissue (e.g., skin, heart or skeletal muscle, aorta), we recommend to extend
More informationPureZOL RNA Isolation Reagent Instruction Manual Catalog #732-6890
PureZOL RNA Isolation Reagent Instruction Manual Catalog #732-6890 For technical support, call your local Bio-Rad office, or in the US, call 1-800-4BIORAD (1-800-424-6723) Table of Contents Section 1 Introduction...1
More informationDNA: A Person s Ultimate Fingerprint
A partnership between the UAB Center for Community Outreach Development and McWane Center DNA: A Person s Ultimate Fingerprint This project is supported by a Science Education Partnership Award (SEPA)
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 informationGentra Puregene Handbook
Third Edition June 2011 Gentra Puregene Handbook For purification of archive-quality DNA from human whole blood bone marrow buffy coat buccal cells body fluids cultured cells tissue mouse tail yeast bacteria
More informationMaxwell 16 Blood DNA Purification System
Technical Manual Maxwell 16 Blood DNA Purification System Caution: Handle cartridges with care; seal edges may be sharp. 2800 Woods Hollow Rd. Madison, WI USA In Vitro Diagnostic Medical Device MDSS GmbH
More informationArcturus PicoPure RNA Isolation Kit. User Guide
Arcturus PicoPure RNA Isolation Kit User Guide For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use. Information in this document is subject to change without notice.
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 informationIn vitro analysis of pri-mirna processing. by Drosha-DGCR8 complex. (Narry Kim s lab)
In vitro analysis of pri-mirna processing by Drosha-DGCR8 complex (Narry Kim s lab) 1-1. Preparation of radiolabeled pri-mirna transcript The RNA substrate for a cropping reaction can be prepared by in
More informationMarmara Üniversitesi Fen-Edebiyat Fakültesi Kimya Bölümü / Biyokimya Anabilim Dalı PURIFICATION AND CHARACTERIZATION OF PROTEINS
EXPERIMENT VI PURIFICATION AND CHARACTERIZATION OF PROTEINS I- Protein isolation and dialysis In order to investigate its structure and properties a protein must be obtained in pure form. Since proteins
More informationNimbleGen DNA Methylation Microarrays and Services
NimbleGen DNA Methylation Microarrays and Services Sample Preparation Instructions Outline This protocol describes the process for preparing samples for NimbleGen DNA Methylation microarrays using the
More informationHow To Get Rid Of Small Dna Fragments
AxyPrep TM Mag FragmentSelect-I Protocol (Fragment Size Selection for Illumina Genome Analyzer and Life Technologies SoLiD) Introduction The AxyPrep Mag FragmentSelect-I purification kit utilizes a unique
More informationVACUETTE RNAgard Blood Tubes BioMaxi Blood RNA Purification Kit. For the collection, preservation and purification of RNA from whole blood
RNAgard Blood System VACUETTE RNAgard Blood Tubes BioMaxi Blood RNA Purification Kit For the collection, preservation and purification of RNA from whole blood Instructions For Use Protocol blood collection,
More informationProcedure for RNA isolation from human muscle or fat
Procedure for RNA isolation from human muscle or fat Reagents, all Rnase free: 20% SDS DEPC-H2O Rnase ZAP 75% EtOH Trizol Chloroform Isopropanol 0.8M NaCitrate/1.2M NaCl TE buffer, ph 7.0 1. Homogenizer-probe
More informationContents. XI. Materials and Equipment Needed But Not Provided 5. DNA Extraction from Small Amount of Tissue 10
Contents I. Overview 1 II. Kit Components 1 III. Storage 2 IV. Intended Use 2 V. Safety Warnings and Precautions 2 VI. Warranty and Liability 2 VII. Technical Assistance 3 VIII. Quality Management 3 IX.
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 informationBacterial Transformation with Green Fluorescent Protein. Table of Contents Fall 2012
Bacterial Transformation with Green Fluorescent Protein pglo Version Table of Contents Bacterial Transformation Introduction..1 Laboratory Exercise...3 Important Laboratory Practices 3 Protocol...... 4
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 informationRiboZol RNA Extraction Reagents
RiboZol RNA Extraction Reagents Code Description Size N580-30ML-SAMPLE Ribozol TM RNA Extraction Reagent 30 ml N580-30ML Ribozol TM RNA Extraction Reagent 30 ml N580-100ML Ribozol TM RNA Extraction Reagent
More informationIsolation of Bacterial DNA
N( EXPERIMENT 19 Isolation of Bacterial DNA Theory To isolate a functional macromolecular component from bacterial cells, you must accomplish three things. First, you must efficiently disrupt the bacterial
More informationMasterPure RNA Purification Kit
Cat. No. MCR85102 Connect with Epicentre on our blog (epicentral.blogspot.com), Facebook (facebook.com/epicentrebio), and Twitter (@EpicentreBio). www.epicentre.com Lit. # 114 6/2012 1 EPILIT114 Rev. A
More informationGenomic DNA Clean & Concentrator Catalog Nos. D4010 & D4011
Page 0 INSTRUCTION MANUAL Catalog Nos. D4010 & D4011 Highlights Quick (5 minute) spin column recovery of large-sized DNA (e.g., genomic, mitochondrial, plasmid (BAC/PAC), viral, phage, (wga)dna, etc.)
More informationSOLIDscript Solid Phase cdna Synthesis Kit Instruction Manual
Toll Free: 866-252-7771 752A Lincoln Blvd. Phone: 732-469-7771 Fax: 732-469-7782 Middlesex, NJ 08846 Web: www.purebiotechllc.com SOLIDscript Solid Phase cdna Synthesis Kit Instruction Manual Product: SOLIDscript
More informationAxyPrep Blood Genomic DNA Maxiprep Kit
AxyPrep Blood Genomic DNA Maxiprep Kit For the purification of high molecular weight genomic DNA from larger volumes of whole blood Kit contents, storage and stability Cat. No. AP-MX-BL-GDNA-10 AP-MX-BL-GDNA-25
More informationStreamlined Purification of Plasmid DNA From Prokaryotic Cultures
Application Note PN 33579 Streamlined Purification of Plasmid DNA From Prokaryotic Cultures Introduction Plasmid DNA has become an essential element of molecular biology and protein research. This invaluable
More informationAffi-Prep Protein A Matrix Instruction Manual
Affi-Prep Protein A Matrix Instruction Manual Catalog Numbers 156-0005 156-0006 Bio-Rad Laboratories, 2000 Alfred Nobel Dr., Hercules, CA 94547 LIT-230 Rev B Table of Contents Section 1 Introduction...1
More informationBacReady TM Multiplex PCR System
BacReady TM Multiplex PCR System Technical Manual No. 0191 Version 10112010 I Description.. 1 II Applications 2 III Key Features.. 2 IV Shipping and Storage. 2 V Simplified Procedures. 2 VI Detailed Experimental
More informationHow To Make A Tri Reagent
TRI Reagent For processing tissues, cells cultured in monolayer or cell pellets Catalog Number T9424 Store at room temperature. TECHNICAL BULLETIN Product Description TRI Reagent is a quick and convenient
More informationBiology 29 Cell Structure and Function Spring, 2009 Springer LABORATORY 2:CHLOROPLASTS AND PHOTOREDUCTION
Biology 29 Cell Structure and Function Spring, 2009 Springer LABORATORY 2:CHLOROPLASTS AND PHOTOREDUCTION In this laboratory we will purify chloroplasts from spinach by differential centrifugation, then
More informationWizard SV Gel and PCR Clean-Up System
TECHNICAL BULLETIN Wizard SV Gel and PCR Clean-Up System Instruc ons for Use of Products A9280, A9281, A9282 and A9285 Revised 12/10 TB308 Wizard SV Gel and PCR Clean-Up System All technical literature
More informationQIAGEN Plasmid Purification Handbook
April 2012 QIAGEN Plasmid Purification Handbook QIAGEN Plasmid Mini, Midi, Maxi, Mega, and Giga Kits For purification of ultrapure, transfection-grade plasmid DNA Sample & Assay Technologies QIAGEN Sample
More informationempcr Amplification Method Manual - Lib-A
GS Junior Titanium Series May 2010 (Rev. April 2011) For life science research only. Not for use in diagnostic procedures. 1. WORKFLOW The emulsion-based clonal amplification (empcr amplification) of a
More informationProtein extraction from Tissues and Cultured Cells using Bioruptor Standard & Plus
Protein extraction from Tissues and Cultured Cells using Bioruptor Standard & Plus Introduction Protein extraction from tissues and cultured cells is the first step for many biochemical and analytical
More informationChIP-IT High Sensitivity
ChIP-IT High Sensitivity (version A4) Catalog No. 53040 Active Motif North America 1914 Palomar Oaks Way, Suite 150 Carlsbad, California 92008, USA Toll free: 877 222 9543 Telephone: 760 431 1263 Fax:
More informationQIAGEN Plasmid Purification Handbook
Second Edition August 2003 QIAGEN Plasmid Purification Handbook QIAGEN Plasmid Midi, Maxi, Mega, and Giga Kits QIAfilter Plasmid Midi, Maxi, Mega, and Giga Kits EndoFree Plasmid Maxi, Mega, and Giga Kits
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 informationGreen Fluorescent Protein (GFP): Genetic Transformation, Synthesis and Purification of the Recombinant Protein
Green Fluorescent Protein (GFP): Genetic Transformation, Synthesis and Purification of the Recombinant Protein INTRODUCTION Green Fluorescent Protein (GFP) is a novel protein produced by the bioluminescent
More informationAssessment of Islet Functional Potency by Glucose Stimulated Insulin Secretion
Page 1 of 5 PURPOSE: To assess the in vitro functional potency of purified islets by measuring the amount of insulin release upon glucose stimulation. SPECIMEN: Purified human Islet preparation. APPLICABLE
More informationChromatin Immunoprecipitation
Chromatin Immunoprecipitation A) Prepare a yeast culture (see the Galactose Induction Protocol for details). 1) Start a small culture (e.g. 2 ml) in YEPD or selective media from a single colony. 2) Spin
More informationChromatin Immunoprecipitation (ChIP)
Chromatin Immunoprecipitation (ChIP) Day 1 A) DNA shearing 1. Samples Dissect tissue (One Mouse OBs) of interest and transfer to an eppendorf containing 0.5 ml of dissecting media (on ice) or PBS but without
More informationKevin Bogart and Justen Andrews. Extraction of Total RNA from Drosophila. CGB Technical Report 2006-10 doi:10.2506/cgbtr-200610
Kevin Bogart and Justen Andrews Extraction of Total RNA from Drosophila CGB Technical Report 2006-10 doi:10.2506/cgbtr-200610 Bogart K and Andrews J. 2006. Extraction of Total RNA from Drosophila. CGB
More informationReadyPrep Protein Extraction Kit (Cytoplasmic/Nuclear)
ReadyPrep Protein Extraction Kit (Cytoplasmic/Nuclear) Instruction Manual Catalog #163-2089 For technical service, call your local Bio-Rad office, or in the US, call 1-800-4BIORAD (1-800-424-6723) Bio-Rad
More informationMouse krebs von den lungen 6 (KL-6) ELISA
KAMIYA BIOMEDICAL COMPANY Mouse krebs von den lungen 6 (KL-6) ELISA For the quantitative determination of mouse KL-6 in serum, plasma, cell culture supernatants, body fluid and tissue homogenate Cat. No.
More informationPureLink HiPure Plasmid Filter Purification Kits
USER GUIDE PureLink HiPure Plasmid Filter Purification Kits For Midi and Maxi preparation of Plasmid DNA Catalog numbers K2100-14, K2100-15, K2100-16, K2100-17, K2100-26, and K2100-27 Revision Date 2 May
More informationBio 3A Lab: DNA Isolation and the Polymerase Chain Reaction
Bio 3A Lab: DNA Isolation and the Polymerase Chain Reaction Objectives Understand the process of DNA isolation Perform DNA isolation using cheek cells Use thermal cycler and Taq polymerase to perform DNA
More informationLAB 7 DNA RESTRICTION for CLONING
BIOTECHNOLOGY I DNA RESTRICTION FOR CLONING LAB 7 DNA RESTRICTION for CLONING STUDENT GUIDE GOALS The goals of this lab are to provide the biotech student with experience in DNA digestion with restriction
More informationBlood Collection and Processing SOP
Brisbane Breast Bank Blood Collection and Processing SOP Breast Pathology Laboratory University of Queensland Centre for Clinical Research Blood Collection We collect 30ml of blood from patients who have
More informationProteoMiner Protein Enrichment Kits
ProteoMiner Protein Enrichment Kits Instruction Manual Catalog # 163-3003 163-3006 163-3007 163-3008 163-3009 163-3010 163-3011 163-3012 For Technical Support, contact your local Bio-Rad office, or in
More informationU.S. Patent No. 9,051,563 and other pending patents. Ver. 1.1.3
INSTRUCTION MANUAL Direct-zol RNA MiniPrep Catalog Nos. R050, R05, R05, & R053 Highlights Quick, spin column purification of high-quality (DNA-free) total RNA directly from TRIzol, TRI Reagent and other
More informationFastLine cell cdna Kit
1. FastLine cell cdna Kit For high-speed preparation of first-strand cdna directly from cultured cells without RNA purification www.tiangen.com RT100701 FastLine cell cdna Kit Cat. no. KR105 Kit Contents
More informationEnzymes: Amylase Activity in Starch-degrading Soil Isolates
Enzymes: Amylase Activity in Starch-degrading Soil Isolates Introduction This week you will continue our theme of industrial microbiologist by characterizing the enzyme activity we selected for (starch
More informationQUANTITATIVE RT-PCR. A = B (1+e) n. A=amplified products, B=input templates, n=cycle number, and e=amplification efficiency.
QUANTITATIVE RT-PCR Application: Quantitative RT-PCR is used to quantify mrna in both relative and absolute terms. It can be applied for the quantification of mrna expressed from endogenous genes, and
More informationMasterPure Complete DNA and RNA Purification Kit
MasterPure Complete DNA and RNA Purification Kit Cat. Nos. MC85200 and MC89010 The MasterPure Complete DNA and RNA Purification Kit provides all of the reagents nec essary to recover nucleic acid from
More informationRapid GST Inclusion Body Solubilization and Renaturation Kit
Product Manual Rapid GST Inclusion Body Solubilization and Renaturation Kit Catalog Number AKR-110 FOR RESEARCH USE ONLY Not for use in diagnostic procedures Introduction Bacteria are widely used for His
More information