pamp, pkan, or pblu?
|
|
- Kristian Miles Owens
- 6 years ago
- Views:
Transcription
1 pamp, pkan, or pblu? Introduction In my final biotechnology lab project, I was given an unidentified plasmid, in which I was required to perform restriction digests using restriction enzymes. A plasmid, however, is a genetic structure in a cell that has the ability to replicate independently of chromosomes. Usually, plasmids are used to analyze DNA, as well as to create proteins. A restriction enzyme is an enzyme that is used to cut and isolate DNA. In order to review the results of our digest, however, I needed to perform a gel electrophoresis. This process allows you to insert your DNA samples into open wells in the gel, and separate the DNA fragments. Then, the gel would be placed into the gel electrophoresis chamber, where the samples will be run using an electrode, with the total distance traveled by the samples ranging between 4-5 centimeters. The goal of this lab was that I would learn how to perform a restriction digests, as well as to understand how to use a gel electrophoresis to separate fragments of DNA of a plasmid. This goal is important because these procedures would help me determine my unidentified plasmid, whether it was pamp, pkan, or pblu, as the title of this report suggests. In order to accomplish this, however, I needed to prepare three solutions into three 1.5 ml microtubes each with an equal amount of the plasmid: the control, the single digest, and the double digest, with the control containing no restriction enzyme. Once these solutions were prepared, I would digest all three of the tubes at 37 degrees Celsius for one hour. Meanwhile, I needed to prepare the gel, so that I could insert the DNA samples into it. However, before adding the samples to the gel, I needed to add 4 microliters of loading dye to the samples so that the DNA fragments would be visible for the photograph. I would then run the gel in the gel electrophoresis chamber to find the sizes of my base pairs. I would then take a picture of the gel, and would later determine the base pair sizes by making a standard curve.
2 Methods Prior to beginning this lab, I performed virtual restriction digests for pamp, pkan, and pblu using the restriction enzymes that I chose on the New England Biolabs website 1 in order to predict my digest results. In order to perform the virtual digests, I gathered the base pair sizes for all 3 plasmids from DNA Learning Center 2. For the double digest, I used the double digest section of the New England Biolabs website 3, in which I tested different combinations of enzymes, which is where I formulated my double digest recipe. If both enzymes could both be digested at the same temperature for the same amount of time, then the combination would work. Once I had performed the virtual digests, I used those numbers as the expected base pair sizes for data analysis. When I was given my plasmid, I found that it had a code of 6410F14. When I began my lab, my lab instructor told me that the starting concentration was 150 ng/ml. When I performed the gel electrophoresis, I used a marker DNA ladder that I would refer to for measurements in order to make my standard curve after the lab. For my restriction enzymes, I used BglI for the single digest, and BamHI & NdeI for the double digest. Then, for the reaction buffer, I used Buffer 3.1. The DNA ladder, the restriction enzymes, and the reaction buffer, all came from New England Bio Labs 1. For my digestion recipe, I calculated that for the single digest for BglI, I would need to digest in a heat block at 37 degrees Celsius for one hour. For my double digest using BamHI & NdeI, I would also need to digest them in a heat block at 37 degrees for an hour. While I waited for an hour, I needed to make my gel for the gel electrophoresis portion of the experiment. I used a 0.8% agarose solution, because we would be focusing on 12,000-8,000 base pairs for the gel. I then needed to measure 0.4 grams of the agarose solution into a flask, then I had to add 5mL of the 10X TAE solution that I prepared earlier, then I needed to bring the volume of the solution to 50 ml using dh 2O. Then, I placed the solution in a microwave, and heated it for about 2-3 minutes in a microwave. Afterward, I added 1 microliter of ethidium bromide to the solution. I poured the solution into a gel gasket, where it would solidify. However, shortly after I poured the solution into the gasket, I would insert a 10-tooth comb into
3 the gel so that I could insert my samples into the gel. Once the gel was prepared and ready to run after I inserted the solutions into the wells. Then, I had to cover the gel, and fill the box, with buffer, in which I did so with a TAE buffer. A month prior to working on this lab, I prepared a 10X TAE Buffer using Tris, Glacial Acidic Acid, and EDTA. A 10X TAE Buffer contains 0.4 moles of Tris with a formula weight of g/mol, 0.2 moles of Glacial Acidic Acid with a formula weight of 17.5 M stock solution, and 0.01 moles of EDTA with a formula weight of g/mol. However, we would only make 250 ml out of 1 L of the Buffer. So, in order to get the proper amounts for the buffer that we would use, I multiplied the moles of each part of the buffer by its molarity, and then multiply it by 0.25, which is the amount of each solution for the buffer. I ended up with g of Tris, 2.86 ml of Glacial Acidic Acid, and g of EDTA. Once we mixed these, we measured the ph level of the buffer using a ph meter, in which we found that the ph level was In this lab, I used 28 ml of the 10X TAE buffer, and then brought the solution to volume 280 ml using about 252 ml of dh 2O. I had to do dilute this in order to make my 1X TAE Buffer. I ran the gel at 140 volts, for approximately minutes. When the DNA solutions ran past the 4 cm mark of the gel tray, I stopped running the gel, and removed it from the gel electrophoresis chamber. Then, I placed it in the UV transilluminator and photographed the gel. To discover the results, I created a standard curve on Microsoft Office Excel. First, I measured the distance traveled by the ladder fragments from the wells of the gel. Next, I recorded the distance, and used the numbers to create a chart with an exponential line, in which I found the equation for the line, which is what I used to help me determine the sizes of my DNA fragments. Then, I measured the distance traveled by all of my DNA fragments from all three samples from the wells of the gel. I plugged in the distance for one fragment in millimeters for x into the equation, in which I would calculate the base pair size of that fragment. The fragment sizes and other data for this lab is in the section below. Results and Conclusions Figure 1
4 Ladder Size (bp) Figure 1 is the image of the gel. Lanes 1: DNA Ladder. Lane 3: Control. Lane 5: Single Digest (BglI). Lane 7: Double Digest (BamHI & NdeI). Lane 9: DNA Ladder Figure 2 Unknown Plasmid y = e x R² = Band migration (mm)
5 Figure 2 shows the standard curve for the gel. The x-axis shows the measured distance traveled of each DNA fragment for the DNA Ladder in millimeters, while the y-axis represents the number of base pairs for the DNA Ladder. The equation at the top right hand corner is the equation for the standard curve that was used to determine the base pair sizes for each fragment of the digests. The R 2 value shows that this graph is reliable because the number is close to 1, in which 1 represents a perfect line. Figure 3 Band Migration Ladder Size (bp) (mm) Ladder Band Ladder Band Ladder Band Ladder Band Ladder Band Ladder Band Ladder Band Ladder Band Ladder Band Control Band Control Band Control Band BglI BglI
6 BglI BglI BglI B & N B & N B & N Figure 3 shows the distances traveled by the digests as well as the base pair sizes for each fragment. I used the equation from Figure 2 to plug in the distance traveled for a fragment for x to find the size of the fragment. Figure 4 BglI (Single Digest) BamHI & Nde I (Double Digest) pamp 3263, 1118, , 1334 pkan 3139, 794, , 235 pblu 2121, 1740, , 2461 Figure 4 shows the expected base pair sizes for pamp, pkan, and pblu for both the single and double digests. As I mentioned earlier in this report, my lab instructor informed me that the initial concentration was 150 ng/ml, and the final concentration would be 250 ng/ml. Figure 1, which is the image of the gel, shows the fragments of each digest. Figure 2 shows the standard curve, along with the equation to
7 determine the base pair sizes of the digests, in which I would use the distance traveled by a fragment in millimeters, and plug that value in for x to find the base pair size for the fragment. The R 2 value shows us that the standard curve is reliable because the value, which is , is close to 1, in which 1 represent a perfect graph. Figure 3 shows the distance traveled in millimeters for the DNA Ladder and the digests, as well as the base pair sizes for every one of them. Figure 4 shows the expected fragment sizes for both the single and double digests for my restriction enzymes that I used for pamp, pkan, and pblu, which I calculated using both the New England Biolabs 1 and the DNA Learning Center 2 websites prior to starting this lab. Unfortunately, when I observed Figure 1, I noticed that the double digest did not digest very well, as the band appeared dark in the picture and the fragments were equivalent to the control, confirming that it did not digest. Thus, it was very hard to determine the plasmid by observing the double digest alone. Although the single digest contained some fragments that appeared as it didn t digest, there were a couple of fragments that I was able to analyze and could determine what my plasmid was based off of those fragments. They were the fragments BglI 3 and BglI 5, as shown on Figure 3, with base pair sizes of 4614 and 1092 respectively. When I observed the expected base pair sizes in Figure 4, I found that the base pair sizes for BglI 3 and BglI 5 were closer to pamp for the single digest, with estimated base pair sizes of 3263 and 1118, which were closer to my actual base pair sizes when compared to the estimated sizes for both pkan and pblu as the base pair sizes for each can be seen in Figure 4 for the single digest. Therefore, after analyzing all of this data, I concluded that my plasmid, with a code of 6410F14, is indeed pamp. References Website: 1 New England Biolabs. Internet: < Website:
8 2 DNA Learning Center. Internet: < Website 3 New England Biolabs. Internet: <
BIOLOGY 163 LABORATORY. RESTRICTION MAPPING OF PLASMID DNA (Revised Fall 2017)
BIOLOGY 163 LABORATORY RESTRICTION MAPPING OF PLASMID DNA (Revised Fall 2017) Physical mapping of genomes is an important part of modern molecular genetics. As it's name implies, physical mapping seeks
More informationLAB 6: Agarose Gel Electrophoresis of Restriction Digested Plasmid DNA
LAB 6: Agarose Gel Electrophoresis of Restriction Digested Plasmid DNA I. Objectives The purpose of today s lab is to learn how to set up and run an agarose gel, separate DNA fragments on the gel, and
More informationDNA RESTRICTION ANALYSIS
DNA RESTRICTION ANALYSIS In this experiment, DNA from the bacteriophage Lambda (48,502 base pairs in length) is cut with a variety of restriction enzymes and the resulting fragments are separated using
More informationLambda (λ) DNA Restriction Digest and Electrophoresis Lab
Lambda (λ) DNA Restriction Digest and Electrophoresis Lab Procedure DAY ONE: restriction digestion Today we will be exposing the lambda DNA to restriction enzymes. For background knowledge, make sure you
More informationGel Electrophoresis and Analysis
6/28/2016 Gel Electrophoresis and Analysis B3 Summer Science Camp at Olympic High School Dr. Jennifer Weller Lab Method: Gel electrophoresis 2 6/28/2016 Electrophoresis: separating molecules in a charged
More informationRFLP ANALYSIS OF DNA LABORATORY
RFLP ANALYSIS OF DNA LABORATORY BIG PICTURE You will be working with an essential research method widely used in genetics, conservation biology, and forensics. The lab is divided into three sections. Part
More informationAnalysis of Precut Lambda DNA. Evaluation copy
Analysis of Precut Lambda DNA Computer 6B Restriction enzymes are a special class of proteins that cut DNA at specific sites and have become an indispensable tool in molecular biology. Restriction enzymes,
More informationCOLLEGE OF THE CANYONS INTRODUCTION TO BIOTECHNOLOGY: CUSTOM LAB
COLLEGE OF THE CANYONS INTRODUCTION TO BIOTECHNOLOGY: CUSTOM LAB GEL ELECTROPHORESIS AND DNA ANALYSIS LAB Version 7-5-12 One of the most basic and frequently used tools of the molecular biologist is electrophoresis.
More informationElectrophoresis 101 Student Worksheet
1 Electrophoresis 101 Student Worksheet Experiment Objective To develop an understanding of electrophoresis principles. To analyze results and to calculate the sizes of unknown charged molecules from given
More informationMOLECULAR GENETICS: TRANSFORMATION AND CLONING adapted by Dr. D. L. Vogelien
Introduction MOLECULAR GENETICS: TRANSFORMATION AND CLONING adapted by Dr. D. L. Vogelien The field of molecular genetics has resulted in a number of practical applications that have been of tremendous
More informationLesson 3 Gel Electrophoresis of Amplified PCR Samples and Staining of Agarose Gels
Lesson 3 Gel Electrophoresis of Amplified PCR Samples and Staining of Agarose Gels What Are You Looking At? Before you analyze your PCR products, let s take a look at the target sequence being explored.
More informationEXPERIMENT GENOMIC DNA ANALYSIS
EXPERIMENT GENOMIC DNA ANALYSIS Population diversity Studies We have 5 species of planarians (3 purchased from Carolina Biologicals, 2 obtained from the Levin lab) andmight have additional species found
More information10 Restriction Analysis of Genomic DNA
10 Restriction Analysis of Genomic DNA Objectives: A) To determine the rough location of restriction sites of an unknown restriction enzyme and B) to use this information to determine the identity of this
More informationThis article reprinted from: Dooley, M. M Restriction endonuclease digestion of a plasmid.
This article reprinted from: Dooley, M. M. 2008. Restriction endonuclease digestion of a plasmid. Pages 389-392, in Tested Studies for Laboratory Teaching, Volume 29 (K.L. Clase, Editor). Proceedings of
More informationHow to Set Up and Run Gel Electrophoresis
How to Set Up and Run Gel Electrophoresis 2 Introduction Gel electrophoresis is a process by which one can separate and analyze various macromolecules based on their size and charge. The process is most
More informationAll-In-One Precast Agarose Gel Electrophoresis Kit (2x9-Well)
All-In-One Precast Agarose Gel Electrophoresis Kit (2x9-Well) Technical Manual No. 0282 Version 03242009 I Description... 1 II Key Features.. 1 III Safety Concerns... 2 IV Kit Contents.... 2 V Storage.....
More informationS-45. What Size Are Your Genes? Edvo-Kit #S-45. Experiment Objective: See page 3 for storage instructions.
Edvo-Kit #S-45 What Size Are Your Genes? S-45 Experiment Objective: The objective of this experiment is to develop an understanding that genetic mutations are inherited from one or both parents. Mutations
More informationMolecular Scissors: Lambda Digest Student Materials
Molecular Scissors: Lambda Digest Student Materials Introduction 2 Pre-Lab Questions. 5 Lab Protocol 6 Data Collection Worksheet. 9 Post-Lab Questions and Analysis.. 10 Plasmid Maps. 13 Last updated: August
More informationNucleic Acid Electrophoresis APPLICATION GUIDE
AGAROSE BUFFERS LADDERS EQUIPMENT Nucleic Acid Electrophoresis APPLICATION GUIDE Reagents: Agarose Thermo Scientific and Fisher Scientific products deliver an end-to-end solution that can meet your most
More informationRestriction Enzyme Analysis of DNA- Student Handout
Restriction Enzyme Analysis of DNA- Student Handout How to set up a restriction enzyme reaction Restriction enzymes (or restriction endonucleases) cleave DNA in a very specific fashion. Type II restriction
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 informationRestriction Enzymes and Lambda DNA
Restriction Enzymes and Lambda DNA Computer 6B Restriction enzymes have become an indispensable tool of molecular researchers over the past fifty years. This unique group of enzymes function as molecular
More informationEZ-Vision DNA Dye as Loading Buffer
EZ-Vision DNA Dye as Loading Buffer Code Description Size N472-SAMPLE EZ-VIsion One, DNA Dye as Loading Buffer, 6X 0.3 ml N650-SAMPLE EZ-Vision Two, DNA Dye as Loading Buffer, 6X 0.3 ml N313-SAMPLE EZ-Vision
More informationDNA Purification for Case Transgene Pronuclear Injection Updated 2/26/08 RM
DNA Purification for Case Transgene Pronuclear Injection Updated 2/26/08 RM Materials: Stratagene StrataPrep DNA Gel Extraction Kit (Catalog #400766 or #400768) http://www.stratagene.com/products/displayproduct.aspx?pid=448
More informationHiPer Random Amplification of Polymorphic DNA (RAPD) Teaching Kit
HiPer Random Amplification of Polymorphic DNA (RAPD) Teaching Kit Product Code: HTBM031 Number of experiments that can be performed: 5 Duration of Experiment: Protocol: 3.5 hours Agarose Gel Electrophoresis:
More informationqpcr Kit, DNA-free Product components 100 rxn 250 rxn Product description
qpcr Kit, DNA-free For the PCR detection and identification of bacterial and fungal DNA using custom primers Product code A8514 Product components 100 rxn 250 rxn A 2.5x mastermix (3 mm MgCl 2 final concentration)
More informationMolecular Scissors: Lambda Digest Teacher Materials
Molecular Scissors: Lambda Digest Teacher Materials Students will conduct a restriction digest of lambda DNA using two unknown enzymes. They will use the results of gel electrophoresis to identify the
More informationAgarose Gel Electrophoresis
Agarose Gel Electrophoresis Gel electrophoresis is a widely used technique for the analysis of nucleic acids and proteins. Agarose gel electrophoresis is routinely used for the preparation and analysis
More informationUsing Single Nucleotide Polymorphism (SNP) to Predict Bitter Tasting Ability
Using Single Nucleotide Polymorphism (SNP) to Predict Bitter Tasting Ability Part II:! Digestion and Analysis of an Amplified Region of the Bitter Taste Receptor TAS2R38 Gene In The Last Lab:! You sampled
More informationRestriction Analysis of Lambda DNA Miriam Golbert, College of the Canyons, Santa Clarita, CO
INTRODUCTION To close the yellow note, click once to select it and then click the box in the upper left corner. To open the note, double click (Mac OS) or right click (Windows) on the note icon. Restriction
More informationImaging Nucleic Acid Gels on the Odyssey Fc Imager
Imaging Nucleic Acid Gels on the Odyssey Fc Imager Developed for: Odyssey Fc Imaging System Published September 2011. The most recent version of this protocol is posted at: https://www.licor.com/bio/support/
More informationHiPer Gel Extraction Teaching Kit (Column Based)
HiPer Gel Extraction Teaching Kit (Column Based) Product Code: HTBM010 Number of experiments that can be performed: 10 Duration of Experiment Agarose Gel Electrophoresis: 1 hour Protocol: 1 hour Agarose
More informationAgarose Gel Electrophoresis Lab
Agarose Gel Electrophoresis ACTIVITY AT A GLANCE Goal: This lab will determine the presence or absence of PCR products and uantify the size (length of the DNA molecule) of the products. Learning Objectives:
More informationLet s Move It! Gel Electrophoresis Using Food Dye Student Guide
Let s Move It! Gel Electrophoresis Using Food Dye Student Guide Purpose This lab explores the principle of electrophoresis, an important technique used in biochemistry and molecular biology. You will:
More informationRestriction Enzyme Mapping
REVISED & UPDATED Edvo-Kit #206 206 Restriction Enzyme Mapping Experiment Objective: In this experiment, students will develop an understanding of plasmid mapping using restriction enzymes. Results are
More informationRAINBOW GELS: AN INTRODUCTION TO ELECTROPHORESIS. STANDARDS 3.1.7, , Westminster College 3.3.7, , 3.3.
RAINBOW GELS: AN INTRODUCTION TO ELECTROPHORESIS STANDARDS 3.1.7, 3.1.10, 3.1.12 Westminster College 3.3.7, 3.3.10, 3.3.12 INTRODUCTION This laboratory will demonstrate the basics of electrophoresis and
More informationPCR Laboratory Exercise
PCR Laboratory Exercise Advance Protocol (updated 1/2018) Introduction Detection of TPA-25 Alu by PCR A Human DNA Fingerprinting Lab Protocol 1994 Cold Spring Harbor Laboratory DNA Learning Center In this
More informationPrinciples and Practice of Agarose Gel Electrophoresis
Edvo-Kit #101 Principles and Practice of Agarose Gel Electrophoresis Experiment Objective: The objective of this experiment is to develop a basic understanding of electrophoretic theory, and to gain "hands-on"
More informationPlasmid Subcloning using low melt ligation
Design Considerations Plasmid Subcloning using low melt ligation General 1) We much prefer directional cloning (since it usually works better and takes less time) and we have found that with the help of
More informationHuman DNA Alu Amplification by Polymerase Chain Reaction (PCR)* Laboratory Procedure
Human DNA Alu Amplification by Polymerase Chain Reaction (PCR)* Laboratory Procedure *Polymerase Chain Reaction is covered by patents owned by Hoffmann-La Roche, Inc. This experiment was adapted from Laboratory
More informationApplications of Biotechnology Electrophoresis lab: (without the DNA) Introduction to micropipetters and electrophoresis equipment
Applications of Biotechnology Electrophoresis lab: (without the DNA) Introduction to micropipetters and electrophoresis equipment Materials- Gather the following items at your table: Eight samples for
More informationIDENTIFICATON OF A TRANSFORMING PLASMID
IDENTIFICATON OF A TRANSFORMING PLASMID Introduction The field of molecular genetics has resulted in a number of practical applications that have been of tremendous benefit to us. One such benefit is the
More informationComparing the Agilent 2100 Bioanalyzer Performance to Traditional DNA Analysis Techniques
Comparing the Agilent 2100 Bioanalyzer Performance to Traditional DNA Analysis Techniques Application Note Author Deborah Vitale Agilent Technologies, Inc. Palo Alto, CA, USA Abstract This Application
More informationAGAROSE GEL ELECTROPHORESIS. Assiut University
AGAROSE GEL ELECTROPHORESIS By Prof. Dr. Asmaa Hussein Prof. of Zoonoses & Director of the MBRU Assiut University The standard method used to separate, identify electrophoresis and purify DNA fragments
More informationAP Biology. Investigation 9: Biotechnology:Restriction Enzyme Analysis of DNA. Investigation 9: Restriction Enzyme Analysis
AP Biology Investigation 9: Biotechnology:Restriction Enzyme Analysis of DNA In this investigation, you will learn how to use restriction Learning Objectives enzymes and gel electrophoresis to create genetic
More informationGeNei TM Gel Extraction Teaching Kit Manual
Teaching Kit Manual Cat No. New Cat No. KT43 106279 KT43A 106300 KT43B 106301 Revision No.: 00280507 CONTENTS Page No. Objective 3 Principle 3 Kit Description 5 Materials Provided 7 Procedure 8 Observation
More informationSynthetic Biology for
Synthetic Biology for Plasmids and DNA Digestion Plasmids Plasmids are small DNA molecules that are separate from chromosomal DNA They are most commonly found as double stranded, circular DNA Typical plasmids
More informationCornell Institute for Biology Teachers
Cornell Institute for Biology Teachers Copyright CIBT This work may be copied by the original recipient from CIBT to provide copies for users working under the direction of the original recipient. All
More informationLecture Four. Molecular Approaches I: Nucleic Acids
Lecture Four. Molecular Approaches I: Nucleic Acids I. Recombinant DNA and Gene Cloning Recombinant DNA is DNA that has been created artificially. DNA from two or more sources is incorporated into a single
More informationGenome Sequence Assembly
Genome Sequence Assembly Learning Goals: Introduce the field of bioinformatics Familiarize the student with performing sequence alignments Understand the assembly process in genome sequencing Introduction:
More informationBC2004 Review Sheet for Lab Exercises 7-11 Spring Semester 2005
BC2004 Review Sheet for Lab Exercises 7-11 Spring Semester 2005 Lab Exercise 7 Drosophila crosses, three weeks Vocabulary: phenotype, genotype, gene, allele, locus (loci), sex chromosomes, autosomes, homozygous,
More informationGuidelines for Using the Sage Science Pippin Pulse Electrophoresis Power Supply System
Please note: the unsupported protocol described herein may not have been validated by Pacific Biosciences and is provided as-is and without any warranty. Use of this protocol is offered to those customers
More informationGene And Cell Technologies
Open Source DNA Ladder Plasmids Gene And Cell Technologies Open-Source DNA Ladder Plasmids can be used to manufacture your own DNA size standards easily and inexpensively. Most people working with DNA
More informationUSDA RiceCAP DNA extraction using DNeasy Plant Mini Kit.
DNA extraction using DNeasy Plant Mini Kit. Preparatory work: 1. If using the kit for the first time, add ethanol to buffer AW and buffer AP3/E to obtain the working solutions. 2. Preheat a water bath
More informationminipcr TM Genes in Space Food Safety Lab: Mars Colony at Risk!
minipcr TM Genes in Space Food Safety Lab: Mars Colony at Risk! An E. coli outbreak affects astronaut food aboard the International Space Station. DNA samples from two food racks are analyzed to determine
More informationFMF NIRCA PROTOCOL STEP 1.
FMF NIRCA PROTOCOL STEP 1. After you have isolated patient s DNA and DNA from a healthy donor (wild type), you perform a nested PCR. The primers used to amplify exon 2 and exon 10 of the mefv gene are
More informationFrequent Difficulties With PFGE (Troubleshooting Tips)
Frequent Difficulties With PFGE (Troubleshooting Tips) 6 th PulseNet Latin America Meeting Buenos Aires, Argentina June 26 th 2008 Efrain M. Ribot, Ph.D. PulseNet Methods Development Laboratory Centers
More information1. For each of these genetic traits which is the dominant allele and which is the recessive allele:
Biology 114 Name: Lab Section Prelab questions for Lab 8 1. For each of these genetic traits which is the dominant allele and which is the recessive allele: being a tongue roller having attached earlobes
More informationManipulation of Purified DNA
Manipulation of Purified DNA To produce the recombinant DNA molecule, the vector, as well as the DNA to be cloned, must be cut at specific points and then joined together in a controlled manner by DNA
More informationLab 3: amplification and isolation of enhancer using PCR & agarose gel extraction
Lab 3: amplification and isolation of enhancer using PCR & Purpose The goal of this lab is to: 1) Dilute your lyophilized primer oligonucleotides to a suitable storage concentration. 2) Design and execute
More informationPTC PCR II: Restriction Enzymes & Gel Electrophoresis
PTC PCR II: Restriction Enzymes & Gel Electrophoresis Objective To apply what we ve learned about genetics, molecular biology, and recombinant DNA to a specific human genetic trait. Background Mammals
More informationLet s Move It! Gel Electrophoresis using Food Dyes Teacher Guide
www.babec.org Let s Move It! Gel Electrophoresis using Food Dyes Teacher Guide Table of Contents Teacher Guide Unit Overview...T1 Inventory Sheet... T2 Background Materials for Teacher Pre-Lab Activity
More informationBiotech CTE Practice Take Home Version A
Biotech CTE Practice Take Home Version A True/False Indicate whether the statement is true or false. 1. Antibiotics can be used to treat a viral infection. Multiple Choice Identify the choice that best
More informationSexing Bovine Preimplantation Embryos by PCR
Sexing Bovine Preimplantation Embryos by PCR Katherine E.M. Hendricks 1, Leydson F. Martins 2, Justin M. Fear 1 and Peter J. Hansen 1 1 Dept. of Animal Sciences, University of Florida and Department of
More informationLAB 1 MOLECULAR GENOTYPING OF Arabidopsis
STUDENT GUIDE LAB 1 MOLECULAR GENOTYPING OF Arabidopsis Information from two NSF sponsored workshops was used in writing this lab: Greenomes: Plant Molecular Genetics and Genomics, taught at the University
More informationMolecular Techniques Third-year Biology
PLANNING Genetics Lab practices Molecular Techniques. Genetics Lab practices protocol. 2015-16 PCR-DIRECTED MUTAGENESIS, MOLECULAR CLONING AND RESTRICTION ANALYSIS Sessions 1 & 2 (2x3 hours): PCR-directed
More informationStudent Manual. Pre-Lab Introduction to DNA Fingerprinting STUDENT MANUAL BACKGROUND
BACKGROUND Pre-Lab Introduction to 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
More informationPreparing Samples for Sequencing Genomic DNA Using the Genomic DNA Sample Prep Oligo Only Kit
Preparing Samples for Sequencing Genomic DNA Using the Genomic DNA Sample Prep Oligo Only Kit Topics 3 Introduction 5 Kit Contents, User-Supplied Reagents, and Equipment 7 Fragment the Genomic DNA 10 Perform
More informationRunOne System Instruction Manual
6 RunOne Electrophoresis System Separate the DNA samples by standard electrophoresis. Prepare a 3x Sybr Green I OR x Sybr Gold Staining Buffer 3x Sybr Green I Staining Buffer: add 5 µl of 0,000x Sybr Green
More informationTruSeq ChIP Sample Preparation
FOR RESEARCH USE ONLY Date: Illumina Kit Description: NOTE Unless familiar with the protocol in the latest version of the TruSeq ChIP Sample Preparation Guide (part # 15023092), new or less experienced
More informationBotanoTech. S c ience N o t ebook. Comparative Plant Genomics Study
BotanoTech S c ience N o t ebook Comparative Plant Genomics Study Table of Contents Introduction & Question... 2 Page Research Phenotype...... 3 Hypothesis. 4 Materials & Methods. 5 Part I Genomic DNA
More informationDescription...1 Components...1 Storage... 1 Technical Information...1 Protocol...2 Examples using the kit...4 Troubleshooting...
QuickClean II Gel Extraction Kit Cat. No. L00418 Technical Manual No. TM0594 Version: 03042011 I II III IV V VI VII VIII Description......1 Components.....1 Storage.... 1 Technical Information....1 Protocol.....2
More informationSAMPLE LITERATURE. Please refer to included weblink for correct version. Colony PCR. Edvo-Kit #323. GFP Transformation Extension:
NOTE: This experiment is designed to work with EDVOTEK Kits 222, 223, or 303. Please refer to page 19 for specifics. Edvo-Kit #323 GFP Transformation Extension: Colony PCR Experiment Objective: In this
More informationE.Z.N.A. Cycle Pure Kit
E.Z.N.A. Cycle Pure Kit D6492-00 5 preps V-spin D6492-01 50 preps V-spin D6492-02 200 preps V-spin D6493-00 5 preps Q-spin D6493-01 50 preps Q-spin D6493-02 200 preps Q-spin March 2017 E.Z.N.A. Cycle Pure
More informationSUNRISE and SUNRISE 96 Gel Electrophoresis Apparatus CAT. SERIES 11068
SUNRISE 12 16 and SUNRISE 96 Gel Electrophoresis Apparatus CAT. SERIES 11068 Table of Contents 1. Notices to Customer... 1 1.1 Important Information... 1 1.2 Warnings... 1 2. Overview... 2 2.1 Description...
More information«ELECTROPHORESIS 2» code EPH02
ELITechGroup S.p.A. C.so Svizzera, 185 10149 Torino ITALY Offices: Tel. +39-011 976 191 Fax +39-011 936 76 11 E. mail: emd.support@elitechgroup.com WEB site: www.elitechgroup.com NOTICE of CHANGE dated
More informationHow Can Pieces of DNA Solve a Puzzle?
Introduction How Can Pieces of DNA Solve a Puzzle? One of the basic tools of modern biotechnology is DNA splicing: cutting DNA and linking it to other DNA molecules. The basic concept behind DNA splicing
More informationXactEdit Cas9 Nuclease with NLS User Manual
XactEdit Cas9 Nuclease with NLS User Manual An RNA-guided recombinant endonuclease for efficient targeted DNA cleavage Catalog Numbers CE1000-50K, CE1000-50, CE1000-250, CE1001-250, CE1001-1000 Table of
More informationRayBio Apoptotic DNA Ladder Extraction Kit
RayBio Apoptotic DNA Ladder Extraction Kit User Manual Version 1.1 March 1, 2016 RayBio Apoptotic DNA Ladder Extraction (Cat#: 68SO-DNAL-S50) RayBiotech, Inc. We Provide You With Excellent Support And
More informationELECTROPHORESIS a es
ELECTROPHORESIS Images DEFINITION Electrophoresis is a procedure for separating a mixture of charged molecules through a stationary material (gel) in an electrical field. It is a powerful tool for separating
More informationEXPERIMENT NINE: SOUTHERN BLOT
EXPERIMENT NINE: SOUTHERN BLOT The objectives of this experiment are to: (1) restrict genomic DNA for Southern blot electrophoresis (2) electrophorese restricted gdna for Southern blotting (3) perform
More informationThe Production of a Recombinant Biotechnology Product. Chapter 8
The Production of a Recombinant Biotechnology Product Chapter 8 Objectives Give a basic overview of genetic engineering. Describe the processes involved in isolating a piece DNA of interest Mass producing
More informationHiPer Yeast Genomic DNA Extraction Teaching Kit
HiPer Yeast Genomic DNA Extraction Teaching Kit Product Code: HTBM013 Number of experiments that can be performed: 10 Duration of Experiment: 3 days Day 1: Revival of Host Day 2: Inoculation of culture
More informationThe GeneEditor TM in vitro Mutagenesis System: Site- Directed Mutagenesis Using Altered Beta-Lactamase Specificity
Promega Notes Magazine Number 62, 1997, p. 02 The GeneEditor TM in vitro Mutagenesis System: Site- Directed Mutagenesis Using Altered Beta-Lactamase Specificity By Christine Andrews and Scott Lesley Promega
More informationGFX PCR DNA and Gel Band Purification Kit
instructions product code: 27-9602-01 GFX PCR DNA and Gel Band Purification Kit Warning For research use only. Not recommended or intended for the diagnosis of disease in humans or animals. Do not use
More informationSickle Cell Gene Detection (DNA-Based)
Edvo-Kit #116 Sickle Cell Gene Detection (DNA-Based) 116 Experiment Objective: In this experiment, students will gain an understanding of the effect of mutations in health and disease, specifi cally as
More informationWow Biolab Gel Electrophoresis
Wow Biolab Free PDF ebook Download: Wow Biolab Download or Read Online ebook wow biolab gel electrophoresis in PDF Format From The Best User Guide Database View the 2-D animation, and answer the following
More informationBacterial 16S rdna PCR Kit Fast (800)
Cat. # RR182A For Research Use Bacterial 16S rdna PCR Kit Fast (800) Product Manual Table of Contents I. Description... 3 II. Components... 3 III. Materials Required but not Provided... 4 IV. Storage...
More informationMicroElute Cycle-Pure Kit
MicroElute Cycle-Pure Kit D6293-00 5 preps D6293-01 50 preps D6293-02 200 preps MicroElute Gel Extraction Kit D6294-00 5 preps D6294-01 50 preps D6294-02 200 preps MicroElute DNA Clean Up Kit D6296-00
More informationBioBrick Formation Step-by-Step Overview: (protocol compiled by Bethany Bruno, Liz Kelly, & Elyse McMillen, last updated 9/24/13)
BioBrick Formation Step-by-Step Overview: (protocol compiled by Bethany Bruno, Liz Kelly, & Elyse McMillen, last updated 9/24/13) As a multistep, complex process, each aspect of biobrick formation takes
More informationLaboratory Exercise 4. Multiplex PCR of Short Tandem Repeats and Vertical Polyacrylamide Gel Electrophoresis.
Laboratory Exercise 4 4 Multiplex PCR of Short Tandem Repeats and Vertical Polyacrylamide Gel Electrophoresis B A C K G R O U N D The human genome contains over 3000 million base pairs, which are distributed
More informationIdentification of Bacterial Species Using Colony PCR
Ouachita Baptist University Scholarly Commons @ Ouachita Honors Theses Carl Goodson Honors Program Spring 2015 Identification of Bacterial Species Using Colony PCR Kaiti Walker Ouachita Baptist University
More informationMonarch DNA Gel Extraction Kit
NUCLEIC ACID PURIFICATION Monarch DNA Gel Extraction Kit Instruction Manual NEB #T1020S/L Version 1.2 3/17 be INSPIRED drive DISCOVERY stay GENUINE This product is intended for research purposes only.
More informationHLA SSP Typing Kits IVD. SSP reagent kit for DNA based HLA typing. Instruction for use. Instruction for use
HLA SSP Typing Kits SSP reagent kit for DNA based HLA typing Product REF Package HLA-A 800 111 24 Tests HLA-B 800 112 24 Tests HLA-DR 800 113 24 Tests HLA-C 800 114 24 Tests HLA-DQ 800 115 24 Tests HLA-
More informationUse of Drosophila Melanogaster as a Model System in the Study of Human Sodium- Dependent Multivitamin Transporter. Michael Brinton BIOL 230W.
Use of Drosophila Melanogaster as a Model System in the Study of Human Sodium- Dependent Multivitamin Transporter Michael Brinton BIOL 230W.001 28 October 2013 TA: Sashi Gollapudi Introduction Many human
More informationHLA SSP Typing Kits. SSP reagent kit for DNA based HLA typing
HLA SSP Typing Kits SSP reagent kit for DNA based HLA typing Product REF Package HLA-A 800 111 24 Tests HLA-B 800 112 24 Tests HLA-DR 800 113 24 Tests HLA-C 800 114 24 Tests HLA-DQ 800 115 24 Tests HLA-
More informationBiotech Term 3 Test. True/False Indicate whether the statement is true or false.
Biotech Term 3 Test True/False Indicate whether the statement is true or false. 1. When you are using a gel to perform electrophoresis, the gel is covered with TAE buffer after you put the DNA in the wells.
More informationSouthern Blot Protocol with Digoxigenin (DIG) probe JAX Mice strain: C57BL/6J Tg(C9orf72_i3)112Lutzy/J
Southern Blot Protocol with Digoxigenin (DIG) probe JAX Mice strain: 023099 C57BL/6J Tg(C9orf72_i3)112Lutzy/J Tail Genomic DNA extraction with QIAGEN DNeasy Blood & Tissue Kit (QIAGEN, cat. no.69504 or
More informationScience Fair Project. Keep Calm and Split DNA Charisma Ware - Carleen McNees - Syracuse Junior High
Science Fair Project Keep Calm and Split DNA Charisma Ware - Carleen McNees - Syracuse Junior High Problem How can you separate DNA when they are to small to see, let alone cut? Hypothesis If I put a group
More informationChapter 11. Restriction mapping. Objectives
Restriction mapping Restriction endonucleases (REs) are part of bacterial defense systems. REs recognize and cleave specific sites in DNA molecules. REs are an indispensable tool in molecular biology for
More information