1 BTEC 4200 Lab 2. Quantifying Bacterial Concentration using a Calibrated Growth Curve Background and References Bacterial concentration can be measured by several methods, all of which you have studied previously in lecture or in lab. In this lab, you will be measuring bacterial concentration by four methods: Turbidity: Turbidity, or light scattering, is measured in a spectrophotometer such as a Spectronic 20. This method has the advantage of being the quickest method at the lab bench. Viable cell count (colony count, or CFU/ml): This is measured by serially diluting a culture, and then spread-plating the dilutions. The concentration of viable cells in the original culture (expressed as colony forming units per ml, or CFU/ml), is calculated from plates counts on the spread plates. Wet mass: A measured volume of a culture is centrifuged to pellet the cells, and then the mass (weight) of the cells is determined. Dry mass: A measured volume of the cells is centrifuged to pellet the cells. The pellet is dried, and the mass (weight) of the dry pellet is determined. For your background, references, and your basic standard procedures you can refer to Unit 8 of the 3340 lab manual (Basic Concepts of the Microbiology Laboratory). You can also reference your textbook (Prescott, Harley, and Klein). Be aware that a thorough and well-written background and procedure is expected as part of your grade for this class. Background: Making a corrected absorbance curve for Escherichia coli In the background section of your lab writeup, you will review the basic theory of the spectrophotometer and how it can be used to measure bacterial concentration (from the 3340 lab manual). In this lab, we are extending our use of turbidity by correcting for deviations from linearity at high absorbance values. You may paraphrase, summarize, and include the following discussion in your background. Central to the idea that the spectrophotometer can be used to measure bacterial concentration is this relationship: Bacterial concentration Absorbance measured with the spectrophotometer This says that the bacterial concentration is directly proportional to the absorbance read on the machine. It means that if the bacterial concentration is doubled, then the number on the machine should double as well. That relationship hold true only up to a particular absorbance value. After that, the culture becomes so turbid that the relationship between bacterial concentration and absorbance begins to deviate from linearity. That means that if you read the absorbance at a value higher that the linear range, the actual bacterial concentration is higher than what you are measuring. This is shown in the data listed in the following table and plotted in the following graph. The data were graphed and analyzed using the Microsoft Excel spreadsheet. 1
2 2 Dilution # Volume of Turbid Culture, ml Volume of Sterile Broth, ml Dilution Factor Measured Absorbance at 425 nm
3 3 Absorbance Correction y = x x R 2 = Absorbance at 425 nm Measured (uncorrected) absorbance Best quadratic fit Dilution Factor In this experiment, a turbid broth culture of E. coli was diluted by varying amounts. The dilution factor equals the volume of the culture in the dilution divided by the total volume of the dilution. For example, 8 ml of the culture mixed with 2 ml of sterile broth gives a dilution factor of 0.8. The 0 dilution was the sterile broth with which the spectrophotometer was blanked. The absorbance at 425 nm of each dilution was measured. If the bacterial concentration were proportional to absorbance for the entire range of concentrations in this dilution series, then the relationship would be linear for the entire range. Instead, linearity is only seen up to an absorbance of approximately 0.5. From this data, it is possible to derive a correction factor to mathematically correct for the deviation from linearity. The correction factor will allow us to make measurements at absorbance values above the linear range, then calculate a corrected absorbance value that is truly proportional to the bacterial concentration. To do this, we first need to fit an equation to the entire data set. A best-fit trendline can be added to the data using Microsoft Excel. In Windows, this is accomplished by right-clicking on the data points and selecting Add Trendline, then selecting Polynomial under Type of trendline. This data fits well to a second order polynomial equation (a good old quadratic equation). The equation for this curve (also shown on the graph) is y = Uncorrected absorbance = A U = x x R 2 = The value R 2 is the correlation coefficient, a statistical term that tells how good the fit is; if the data fits the curve perfectly, then the R 2 value is 1. An R 2 value of indicates an excellent fit, as the picture of the graph also shows.
4 4 The next step in deriving our correction factor is to get a second equation for the linear part of the graph. The linear equation will serve as the basis of our corrected absorbance value. To do this, we plot the linear portion of the data on a separate graph. Absorbance Correction y = x R 2 = Absorbance at 425 nm Measured (uncorrected) absorbance Best linear fit Dilution Factor The data are plotted in Microsoft Excel and a trendline is determined. In this case, we determine the best linear trendline to the data as the basis for our corrected absorbance: y = Corrected absorbance = A C = x R 2 = say): Now we have two equations (a set of simultaneous equations, as the mathematicians A U = x x A C = x What we do now is to take the first equation and solve for x as a function of A U. Then, we plug x into the second equation and we have our correction formula.
5 5 To solve the first equation, we use the quadratic formula L: A U = x x 0 = x x A U ± x = (1.5305) 2 2( 0.449) 4( 0.449)( A U ) ± x = x = ± A A U U Like most calculations with a quadratic equation, we have to figure out whether the plus or the minus is the correct sign in front of the radical. This is easily done by taking a measured A U from the data at a specific x value (dilution factor), plugging the A U into the equation, doing the calculations with the plus and the minus signs, and seeing which one gives the correct x value (dilution factor). At a dilution factor (x value) of 0.1, the measured A U was Entering the value for A U into the equation and calculating possible x values using the plus and minus signs, we get: Using plus sign: x = (not close to 0.1) Using the minus sign: x = (almost equal to 0.1). Therefore, the correct sign in our calculation is the minus sign. x = A U Having solved for x as a function of A U, now we can enter this into the second of the two equations to get our correction formula. A C = x A C = A U A = C A U The graph on the next page shows the data set of measured absorbance values (A U ) plotted together with the corrected absorbance values determined from the formula.
6 6 Absorbance Correction y = 1.381x R 2 = Absorbance at 425 nm y = x x R 2 = Measured (uncorrected) absorbance. Corrected absorbance Best quadratic fit (uncorrected absorbance) 0.2 Best linear fit (corrected absorbance) Dilution Factor There are several points to note in this analysis. In the range of linearity, there is no significant difference between the measured (uncorrected) absorbance values and the corrected absorbance values, as we would expect. The corrected absorbance values give an excellent linear fit (R 2 = 0.999) with the same slope as the linear region of the uncorrected absorbance values, again as we would expect. The formula is only valid for calculations on uncorrected absorbance values up to 1.3. At A U values just over 1.3, the term under the radical becomes negative, so an A C value cannot be calculated. This is not an issue, because a broth culture will rarely reach an absorbance as high as 1.3. Furthermore, an absorbance of 1.3 represents a percent transmittance of about 5%; in other words, a suspension so turbid that it is almost opaque. If one had to make an optical measurement of such a suspension, there would be no choice but to dilute the suspension, measure the absorbance (and correct if necessary), and multiply the absorbance times the dilution factor. The correction formula may be different for different bacterial species, or for the same species grown in different media or grown under different conditions. The data presented above were collected on E. coli grown in Minimal Davis broth medium. In this laboratory exercise, you will be determining a similar correction factor for E. coli grown in tryptic soy broth.
7 7 Procedure: Making a corrected absorbance curve for Escherichia coli Note: This may be used as written, except that you should include the table with the volumes in your lab notebook. I just didn t want to have to put it down twice. 1. Be sure to wear gloves during this procedure, to avoid getting culture on your hands while transferring liquid from tubes. 2. Turn on a Spectronic 20 spectrophotometer and set the wavelength to 425 nm. Let it warm up for at least 15 min. 3. Obtain a turbid broth culture of E. coli, tryptic soy broth (TSB), 21 clean culture tubes (they do not need to be sterile), a clean Spectronic 20 tube, three 10-ml pipets, and a pipump. 4. Label the culture tubes Using different pipets, add turbid E. coli culture and TSB to each tube in the volumes shown in the table on page Mix each tube thoroughly by vortexing for 1 2 sec with a vortex mixer. 7. With no tube in the spectrophotometer, set the 0%T setting. 8. Transfer 3 4 ml of broth from tube 21 (TSB with no E. coli) to the Spectronic 20 tube, carefully wipe the outside of the tube with a kimwipe, place the tube in the machine, and set the 100%T setting (blank the spectrophotometer). Switch the spectrophotometer setting to Absorbance. 9. Remove the Spectronic tube and pour the broth back into tube 21. Carefully drain the last few drops from the lip of the Spectronic tube into a kimwipe. Put the contaminated kimwipe in the biohazard bag. 10. Starting with tube #20 and going backwards to tube #1, transfer 3 4 ml of each dilution to the Spectronic tube, wipe the tube with a kimwipe, place the tube in the spectrophotometer, and read the absorbance. After each tube, pour the broth back into its numbered tube and drain the last few drops into a kimwipe, as described earlier. 11. Place caps on the culture tubes, remove the labels, and place the tubes in the proper area for cleanup. 12. Transfer the data to a Microsoft Excel spreadsheet. The spreadsheet should contain the following columns: volume of E. coli culture, volume of broth, dilution factor, and measured absorbance. 13. In the spreadsheet, create an X-Y scatter plot of dilution factor vs absorbance. Note: All charts must be added as separate sheets in the workbook, not pasted into the worksheet with the data columns. 14. Add a second order polynomial trendline to the plot, and set the options on the trendline to show the equation and the correlation coefficient. 15. From the plot, identify the range in which absorbance is proportional to bacterial concentration. 16. Create a second plot, plotting only the linear range of dilution factor vs absorbance. 17. Add a linear trendline to the second plot, and set the options for the trendline to show the
8 8 equation and the correlation coefficient. 18. Using the equations from the first and second plot, derive a correction formula to correct for linearity at high absorbance readings. 19. Add another column to the spreadsheet: Corrected absorbance. The values in the Corrected absorbance column are calculated from the Measured absorbance column using the correction formula, entered in Microsoft Excel notation. For example, if the correction formula is A = C A U then the formula entered in the spreadsheet cell is = (1.5375*SQRT( (1.796*D7))) where D7 is the reference to the cell containing the Measured absorbance (A U ) value. 20. Create a third plot with two data series: dilution factor vs. measured absorbance, and dilution factor vs. corrected absorbance. The easiest way to do this is to copy the first chart (by right clicking on its tab), going to the copy of the chart, selecting the data points (by right clicking on them), selecting Source data Series, clicking Add series, and setting the Y-value ranges on the new series to correspond to the Corrected absorbance column. 21. On the third plot, add a second order polynomial trendline to the dilution factor vs measured absorbance plot, and a linear trendline to the dilution factor vs corrected absorbance plot.
9 9 Background: Calibrating absorbance measurements with viable cell counts, wet mass, and dry mass measurement In the background section of your laboratory writeup, you will review the basic concepts of the viable cell count technique, given in your 3340 laboratory manual (Basic Concepts of the Microbiology Laboratory), your textbook, and the lecture notes from this class. In this lab, we will determine the viable cell count (CFU/ml) of a turbid broth culture of E. coli, making the following modifications to the procedure from the 3340 lab manual. We will measure the absorbance of the E. coli culture before making the serial dilutions and plates. The measured absorbance will be corrected using the correction formula. After determining the CFU/ml from the plate counts, we will have established a calibration factor to convert absorbance (A C ) into CFU/ml. In two separate centrifuge tubes, we will centrifuge 10 ml of the turbid culture to pellet the cells. After discarding the broth, we will determine the wet mass (wet weight) of the cells, then we will allow the cells dry out to determine the dry mass. After determining the CFU/ml from the plate counts, we will have established a calibration factor to convert absorbance (A C ) into wet mass (gm wet cells/ml) or dry mass (gm dry cells/ml). We will plate duplicate plates of each dilution. Plates will be counted with valid counts in the range of colonies per plate, according to the FDA Methods of Bacteriological Analysis. Also, we will round the counts to two significant figures after making our calculations. The CFU/ml in the broth culture will be determined by the formula CFU C = ml [(1*n ) + (0.1* n2) ]*d1*v Where: C = Sum of all colonies on all plates between n 1 = number of plates counted at dilution 1 (least diluted plate counted) n 2 = number of plates counted at dilution 2 (dilution 2 = 0.1 of dilution 1) d 1 = dilution factor of dilution 1 V= Volume plated per plate
10 10 Procedure: Calibrating absorbance measurements with viable cell counts, wet mass, and dry mass measurement Materials 1. Obtain a turbid broth culture of E. coli, 16 sterile culture tubes with caps, sterile tryptic soy broth, 32 tryptic soy agar plates, two Sorvall centrifuge tubes, a Spectronic 20 tube, three sterile 10-ml pipets, a sterile 1-ml pipet, pi-pumps, vortex mixer, and 16 sterile plastic disposable inoculating loops. Determining the corrected absorbance of the E. coli culture 2. Warm up a Spectronic 20 spectrophotometer for at least 15 min and set the wavelength to 425 nm. With no tube in the machine, set 0%T. Blank the spectrophotometer (100%T) with sterile tryptic soy broth. 3. Transfer 3 4 ml of the E. coli culture into the Spectronic 20 tube, and measure the absorbance. Record the absorbance in your lab notebook. 4. Discard the used culture from the Spectronic tube into a waste container, and repeat the procedure to get a replicate reading. Record the absorbance, and average the two readings together to get the uncorrected absorbance (A U ) for the culture. 5. Using the correction formula determined in the earlier procedure, calculate a corrected absorbance (A C ) value for the culture. Wet mass and dry mass measurements 6. With a laboratory marker, label the Sorvall tubes 1 and 2. Note: the tubes may have already been permanently numbered; if so, just use the numbers on the tubes and make note of the numbers in your lab book. 7. Weigh each of the Sorvall tubes, and record their weights in your lab notebook. 8. Carefully measure 10 ml of the E. coli culture into each Sorvall tube. 9. Centrifuge the tubes at 5000 rpm for 10 min in the Sorvall RC-5 centrifuge. 10. Decant the supernatant into the waste container. Carefully blot the last few drops from the lip of the tube with a kimwipe. Discard the kimwipe in the biohazard bag. 11. Weigh the tubes with the wet cell pellets. Subtract the weight of tube+pellet minus tube alone to get the weight of the wet cell pellet. 12. Divide the wet weight of cells by 10 ml to get the concentration of E. coli in the broth culture, expressed in gm wet cells/ml. Do this for each tube, and average the two values. 13. Divide the value obtained in step 12 (gm wet cells/ml) by the value obtained in step 5 (A C ) to obtain the calibration factor to convert A C into gm wet cells/ml. 14. Place the open tubes, in a test tube rack, in a biosafety hood with the UV lamp turned on. Let the tubes sit opened for 2 days to dry out in the hood. 15. Weigh the tubes with the dry cell pellets. Subtract the weight of tube+pellet minus tube alone to get the weight of the dry cell pellet. 16. Divide the dry weight of cells by 10 ml to get the concentration of E. coli in the broth culture, expressed in gm dry cells/ml. Do this for each tube, and average the two values. 17. Divide the value obtained in step 16 (gm dry cells/ml) by the value obtained in step 5 (A C ) to obtain the calibration factor to convert A C into gm dry cells/ml.
11 11 Viable cell count measurements 18. Label the sterile culture tubes Carefully add 9 ml of TSB to each culture tube. 20. Label two TSA plates 10-1, two 10-2, etc., through Using a 1-ml pipet, transfer 1 ml of the E. coli culture to the 10-1 tube. Do not discard the pipet. 22. Mix the dilution thoroughly by vortexing 1 2 sec. 23. Rinse the pipet 3 4 times in the 10-1 dilution, then carefully withdraw 1 ml of the 10-1 dilution with the pipet and transfer it to the 10-2 tube. Note: the purpose of the pipet rinsing step is to completely coat the inside of the pipet with the current dilution so the same pipet can be used for each dilution in the series with negligible error. Otherwise, the inside of the pipet would be coated with the previous dilution, which is tenfold higher than the dilution we are pipetting. 24. Mix the dilution, rinse the pipet, and continue the procedure until all the serial dilutions have been made. 25. Discard the pipet in the appropriate used glass container. 26. Using a new 1-ml pipet and beginning with the dilution, pipet 0.1 ml of the dilution onto an appropriately labeled TSA plate (do not discard the pipet), and immediately spread the drop with a plate spreader formed from a plastic inoculating loop. Note: the spreader can be made by bending the needle portion of the loop into an L shape in the sterile lid of the petri dish. 27. Repeat this procedure with the duplicate plate. Do not discard the pipet. Note: the reason for starting with the dilution is so the same pipet can be used for all the dilutions. 28. Rinse the pipet 3 4 times with the dilution, and repeat the plating procedure (steps 26 27). 29. Incubate the plates at 37 C for 48 hr. 30. Count the colonies on plates having colonies. 31. Determine the CFU/ml in the broth culture by the formula CFU C = ml [(1*n ) + (0.1* n2) ]*d1*v where: C = Sum of all colonies on all plates between n 1 = number of plates counted at dilution 1 (least diluted plate counted) n 2 = number of plates counted at dilution 2 (dilution 2 = 0.1 of dilution 1) d 1 = dilution factor of dilution 1 V= Volume plated per plate 32. Divide the value obtained in step 31 (CFU/ml) by the value obtained in step 5 (A C ) to obtain the calibration factor to convert A C into CFU/ml.
Spreadsheets and Laboratory Data Analysis: Excel 2003 Version (Excel 2007 is only slightly different) Spreadsheets are computer programs that allow the user to enter and manipulate numbers. They are capable
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
Calibration of Glassware Introduction Glassware is commonly calibrated using a liquid of known, specific density, and an analytical balance. The procedure is to determine the mass of liquid the glassware
Determining the Quantity of Iron in a Vitamin Tablet Computer 34 As biochemical research becomes more sophisticated, we are learning more about the role of metallic elements in the human body. For example,
Test Method for the Continuous Reduction of Bacterial Contamination on Copper Alloy Surfaces Test Organisms: Staphylococcus aureus (ATCC 6538) Enterobacter aerogenes (ATCC 13048) Pseudomonas aeruginosa
Bacterial Transformation with Green Fluorescent Protein pglo Version Table of Contents Bacterial Transformation Introduction..1 Laboratory Exercise...3 Important Laboratory Practices 3 Protocol...... 4
Transformation of the bacterium E. coli using a gene for Green Fluorescent Protein Background In molecular biology, transformation refers to a form of genetic exchange in which the genetic material carried
Induction of Enzyme Activity in Bacteria:The Lac Operon Preparation for Laboratory: Web Tutorial - Lac Operon - submit questions I. Background: For the last week you explored the functioning of the enzyme
EXCEL Tutorial: How to use EXCEL for Graphs and Calculations. Excel is powerful tool and can make your life easier if you are proficient in using it. You will need to use Excel to complete most of your
Transformation Kit BACTERIAL TRANSFORMATION: GREEN FLUORESCENT PROTEIN Partnership for Biotechnology and Genomics Education Barbara Soots Linda Curro Education Coordinator University of California Davis
Lab Exercise 3: Media, incubation, and aseptic technique Objectives 1. Compare the different types of media. 2. Describe the different formats of media, plate, tube etc. 3. Explain how to sterilize it,
Introduction Gloves and laboratory coats Small tools Specific clean-up and housekeeping procedures are used to help protect evidence samples from conditions and agents that might serve to destroy, deteriorate,
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
Microsoft Excel Tutorial by Dr. James E. Parks Department of Physics and Astronomy 401 Nielsen Physics Building The University of Tennessee Knoxville, Tennessee 37996-1200 Copyright August, 2000 by James
Rat Creatine Kinase MB isoenzyme,ck-mb ELISA Kit Catalog No: E0479r 96 Tests Operating instructions www.eiaab.com FOR RESEARCH USE ONLY; NOT FOR THERAPEUTIC OR DIAGNOSTIC APPLICATIONS! PLEASE READ THROUGH
Calibration and Linear Regression Analysis: A Self-Guided Tutorial Part 1 Instrumental Analysis with Excel: The Basics CHM314 Instrumental Analysis Department of Chemistry, University of Toronto Dr. D.
Case File 9 Killer Cup of Coffee: Using colorimetry to determine concentration of a poison Determine the concentration of cyanide in the solution. A Killer Cup of Coffee? SOUTH PAINTER, Tuesday: It was
Send To: 1X220 Ms. Laura Marshall AcornVac, Inc. 13818 Oaks Avenue Chino CA 91710 United States Facility: 1X221 AcornVac, Inc. 13818 Oaks Avenue Chino CA 91710 United States Result: COMPLETE Report Date:
HARVESTING AND CRYOPRESERVATION OF HUMAN EMBRYONIC STEM CELLS (hescs) OBJECTIVE: can be cryopreserved in a liquid nitrogen (LN 2 ) freezer for long-term storage. This Standard Operating Procedure (SOP)
THE LABORATORY NOTEBOOK In scientific work keeping a permanent record of all raw data, observations, calculations, et cetera obtained during an experiment is important. Therefore, a student must become
1 of 8 The quality assurance plan as presented in the Quality Manual should assure that constant and consistent test conditions are met and verified and should be tailored to the laboratory s activities.
CHEM 311L Quantitative Analysis Laboratory Revision 2.3 Calibration of Volumetric Glassware In this laboratory exercise, we will calibrate the three types of glassware typically used by an analytical chemist;
National Food Safety Standard of the People s Republic of China GB4789.2-2010 National Food Safety Standard Food microbiological examination: Aerobic plate count Issued by 2010-03-26 Implemented by 2010-06-01
BLS-4 Biosafety Level 2 (BSL-2) Safety Guidelines BSL-3 BSL-2 BSL-1 BSL-2 builds upon BSL-1. If you work in a lab that is designated a BSL-2, the microbes used pose moderate hazards to laboratory staff
1 ATOMIC ABSORTION SPECTROSCOPY: rev. 4/2011 ANALYSIS OF COPPER IN FOOD AND VITAMINS Buck Scientific Atomic Absorption Spectrophotometer, Model 200 Atomic absorption spectroscopy (AAS) has for many years
Experiment 0 Introduction to Data Analysis Using an Excel Spreadsheet I. Purpose The purpose of this introductory lab is to teach you a few basic things about how to use an EXCEL 2010 spreadsheet to do
OVERVIEW Prevention is the most important part of any spill management plan. Be sure to read and understand standard operating procedures (SOP s) and protocols for safe manipulation of biohazards before
Use of Micropipettes Prior to lab you should understand: The function of micropipettes in the laboratory Basic parts of micropipette What volumes are measured with P, P and P1 micopipettors How to read
EXERCISE 2 Name How do scientists prepare solutions with specific concentrations of solutes? Objectives After completing this exercise, you should be able to: define and correctly use the following terms:
I. Purpose Biohazardous material usage on the Scott & White campus is regulated by the Scott & White Institutional Biosafety Committee (IBC). Those investigators choosing to perform research with biohazardous
A Guide to Using Excel in Physics Lab Excel has the potential to be a very useful program that will save you lots of time. Excel is especially useful for making repetitious calculations on large data sets.
KAMIYA BIOMEDICAL COMPANY Mouse Creatine Kinase MB isoenzyme (CKMB) ELISA For the quantitative determination of mouse CKMB in serum, plasma, cell culture fluid and other biological fluids Cat. No. KT-57681
Introduction INTRODUCTION The Microbiology Prep Room is located in 531A Life Sciences Building. The telephone number is 372-8609. It is open from 7:30 a.m. to 4:30 p.m. during the fall and spring semesters.
How uch Phosphate is the Body Being Exposed to During a Lifetime by Showering? Lab 5: Quantitative Analysis- Phosphates in Water By: A Generous Student LBS 171L Section 9 TA: Dana October 7, 005 [Note:
Epinephrine/Norepinephrine ELISA Kit Catalog Number KA3767 96 assays Version: 01 Intended for research use only www.abnova.com Table of Contents Introduction... 3 Intended Use... 3 Principle of the Assay...
Home Contacts Order Catalog Support Search Alphabetical Index Numerical Index Restriction Endonucleases Modifying Enzymes PCR Kits Markers Nucleic Acids Nucleotides & Oligonucleotides Media Transfection
USER GUIDE One Shot TOP10 Competent Cells Catalog Numbers C4040-10, C4040-03, C4040-06, C4040-50, and C4040-52 Document Part Number 280126 Publication Number MAN0000633 Revision A.0 For Research Use Only.
Excel Tutorial Below is a very brief tutorial on the basic capabilities of Excel. Refer to the Excel help files for more information. Working with Data Entering and Formatting Data Before entering data
Radiation Sciences Nuclear Medicine Technology Program Hot Lab Log Book (Material) Table of Content Receiving a Radioactive Package Constancy Test Daily Survey Weekly Wipes Accuracy Test Linearity Geometric
DNA Analyst Training Laboratory Training Manual Protocol 2.02 Clean Technique This laboratory protocol (or part thereof) has been provided as an example of a laboratory SOP, courtesy of the Illinois State
Yale University Office of Environmental Health & Safety Biosafety Spill Response Guide Office of Environmental Health & Safety 135 College Street, 1 st Floor, New Haven, CT 06510 Telephone: 203-785-3550
Rat creatine kinase MM isoenzyme (CK-MM) ELISA Kit Catalog Number. CSB-E14405r For the quantitative determination of rat creatine kinase MM isoenzyme (CK-MM) concentrations in serum, plasma, tissue homogenates.
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
Tutorial 2: Using Excel in Data Analysis This tutorial guide addresses several issues particularly relevant in the context of the level 1 Physics lab sessions at Durham: organising your work sheet neatly,
Entering and Formatting Data Using Microsoft Excel to Plot and Analyze Kinetic Data Open Excel. Set up the spreadsheet page (Sheet 1) so that anyone who reads it will understand the page (Figure 1). Type
INTRODUCTION Phosphates (ortho- and total) Phosphorus is an essential nutrient for all aquatic plants and algae. Only a very small amount is needed, however, so an excess of phosphorus can easily occur.
Engineering Problem Solving and Excel EGN 1006 Introduction to Engineering Mathematical Solution Procedures Commonly Used in Engineering Analysis Data Analysis Techniques (Statistics) Curve Fitting techniques
Revision No.: ZJ0002 Issue Date: Aug 7 th, 2008 HBV Quantitative Real Time PCR Kit Cat. No.: HD-0002-01 For Use with LightCycler 1.0/LightCycler2.0/LightCycler480 (Roche) Real Time PCR Systems (Pls ignore
A Guide to Managing Your Biological Waste at the University at Albany Section 1 - What you need to know: Definition: "Regulated Medical Waste (RMW) shall mean any of the following waste which is generated
Carnegie Mellon University s Policy and Procedures for Recombinant and Synthetic Nucleic Acid Materials Spills Background In accordance with Section IV-B-2-b-(6) of the NIH Guidelines for Research Involving
UNIT PLAN: EXPONENTIAL AND LOGARITHMIC FUNCTIONS Summary: This unit plan covers the basics of exponential and logarithmic functions in about 6 days of class. It is intended for an Algebra II class. The
PM 2.5 Nylon Filter Cleaning Date: August 25,2009 Page I of8 Standard Operating Procedure for Cleaning Nylon Filters Used for the Collection of PM 2. 5 Material Environmental and Industrial Sciences Division
Page: 1 of 7 1. Purpose: 1.1. To describe the procedures to be used when dealing with chemical or microbiological spills. 2. Equipment: 2.1. Spill Kit 2.2. Miscellaneous items as listed 3. Materials: 3.1.
Tutorial on Using Excel Solver to Analyze Spin-Lattice Relaxation Time Data In the measurement of the Spin-Lattice Relaxation time T 1, a 180 o pulse is followed after a delay time of t with a 90 o pulse,
Mouse Insulin ELISA For the quantitative determination of insulin in mouse serum and plasma Please read carefully due to Critical Changes, e.g., Calculation of Results. For Research Use Only. Not For Use
Instrument Adaptation The attached instrument adaptation has been prepared and validated by the reagent manufacturer, Hyphen-Biomed. Instrument Siemens BCS-XP Product BIOPHEN Heparin LRT Analyte Rivaroxaban
Page 1 Data representation and analysis in Excel Let s Get Started! This course will teach you how to analyze data and make charts in Excel so that the data may be represented in a visual way that reflects
Quality Assurance/Quality Control in Acid Deposition Monitoring Acid Deposition and Oxidant Research Center (Network Center of EANET) Introduction Wet deposition data are used for -assessments of spatial
Response to Biological Spills in the Laboratory (Intentional or Accidental) Exposure Management For splash to eyes, mucous membranes, or broken area of the skin Irrigate eyes with clean water, saline or
Generating ABI PRISM 7700 Standard Curve Plots in a Spreadsheet Program Overview The goal of this tutorial is to demonstrate the procedure through which analyzed data generated within an ABI PRISM 7700
Cautions: 6 M hydrochloric acid is corrosive. Purpose: To colorimetrically determine the mass of iron present in commercial vitamin tablets using a prepared calibration curve. Introduction: Iron is considered
1 Lab 2 Biochemistry Learning Objectives The lab has the following learning objectives. Investigate the role of double bonding in fatty acids, through models. Developing a calibration curve for a Benedict
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.
Chestnut Leaf DNA Extraction Protocol Introduction: we will extract the nucleic acid from leaf tissue by grinding it in a reducing medium (the beta-mercaptoethanol chemical is a reducing agent, it smells
BIOLOGICAL SPILL KIT IN A 5 GALLON BUCKET WITH LID Spill response and cleanup procedures (SOP) 1 Notepad 1 Pen 6 Business cards 1 Permanent marker 1 trash bag 6 Biohazard stickers 1 roll duct tape 1 roll
Name: Date: Understanding Analytical Chemistry (Weighing, Mixing, Measuring and Evaluating) High School Environmental Science AP Module 1 Environmental Lab NGSSS Big Ideas: This module is a laboratory-based
AEROBIC RESPIRATION LAB DO 2.CALC From Biology with Calculators, Vernier Software & Technology, 2000. INTRODUCTION Aerobic cellular respiration is the process of converting the chemical energy of organic
Recommended Procedures for Use of YSI Water Quality Monitoring Instruments during Oil Spills Overview: This is a guidance document that is intended for users of YSI 6-Series sondes and handheld instruments
Infectious Waste Management Plan Infectious Waste Management Plan USC Health & Safety Programs Unit 777-5269 POLICY: A. In keeping with the University of South Carolina's policy of providing protection
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
Protocol for Disinfection of Cell Culture and Tissue Culture in Media: Location: Hickory Hall 001 Director: Dr. Guido Verbeck DECONTAMINATION OF CELL CULTURE WASTE Cell culture has become a common laboratory
Estimating a market model: Step-by-step Prepared by Pamela Peterson Drake Florida Atlantic University The purpose of this document is to guide you through the process of estimating a market model for the
FOR IN VITRO AND RESEARCH USE ONLY NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES SEB268Mu 96 Tests Enzyme-linked Immunosorbent Assay Kit For Phenol Sulfotransferase (PST) Organism Species: Mus musculus
QSM Approval: Coating and Extraction of Honeycomb Denuders 1 INTRODUCTION 1.1 The following procedures are used for the coating of honeycomb denuders with citric acid and sodium carbonate solutions, and
Guide to using the Bio Rad CFX96 Real Time PCR Machine Kyle Dobbs and Peter Hansen Table of Contents Overview..3 Setup Reaction Guidelines 4 Starting up the Software 5 Setup Protocol on Software 6 Setup
Exploring Relationships between Highest Level of Education and Income using Corel Quattro Pro Created by Michael Lieff (firstname.lastname@example.org) Faculty of Education, Queen s University While on practicum at Statistics
Provider York County School Division Course Syllabus URL http://yorkcountyschools.org/virtuallearning/coursecatalog.aspx Course Title Algebra I AB Last Updated 2010 - A.1 The student will represent verbal
Instructions for use PCR KIT FOR EXTRACTION OF RNA AND REAL TIME PCR DETECTION KIT FOR HEPATITIS C VIRUS RNA Research Use Only Qualitative Uni Format VBD0798 48 tests valid from: December 2013 Rev11122013
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
Lab Safety and Standard Operating Procedures Faculty of Dentistry And School of Biomedical Engineering Introduction It is the requirement that students working in research laboratories at Dalhousie have