4. Follow-up questions Q1. Why SYBRSafe is needed in DNA gel electrophoresis? SYBRSafe is a dye for DNA staining to determine the presence of DNA fragment in gel electrophoresis. The SYBRSafe performs like ethidium bromide that binds to DNA fragment. SYBRSafe is excited by UV or strong light (e.g. blue light) and emits fluorescent signal which is proportional to the concentration of DNA in the gel (Thermofisher Scientific‚ 2011). In this experiment‚ SYBRSafe is used for the detection of mutant
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Beta-Galactosidase and Western Blot 0 1. EMG 9 and EMG 26 contain strain _lac-_(I- Z+ Y+) and strain _lac -_ (I+ Z- Y-)respectively.Three genes huddled together on the chromosome are required for two strains of _E.coli_ to utilize lactose.Consisting of three genes‚ namely‚ _lacZ_‚ _lacY_ and _lacA_‚ the _lac_ operon orderly handles these genes to code specific enzymes necessary for the metabolism of lactose. The genes _lacZ_‚ _lacY_ and _lacI_ would code for beta-galactosidase‚ galactosidase permease
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Study Guide for the BME5 Midterm The midterm will cover information from lectures 1-4. Subject areas that are fair game for the test are listed below. I may not use every single item in the exam. Note that the level of detail required is that which was covered in lecture. Only information covered in lecture will be on the tests (includes info in figures and tables). Lecture 1 Processes that utilize recombinant DNA technology (and molecular processes that don’t) Basic science; what it is‚
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com/wiki/-/wiki/Main/Applications+of+Cloning http://c2d.osdd.net/home/cep/intro http://www.ncbi.nlm.nih.gov/books/NBK21498/ http://www.nature.com/scitable/topicpage/the-biotechnology-revolution-pcr-and-the-use-553 http://www.mrothery.co.uk/genetech/genetechnotes.htm#Electrophoresis
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eluate imidazole‚ nxt we do size exclusion which separates small from big‚big comes off first the imidazole captures it in the column next we do spectrophotometry. Then SDS-PAGE electrophoresis‚ size purity‚enzyme test does its function. Finally we did DHFR enzymatic assay. In the SDS-PAGE electrophoresis we load the gel up with Laemmli buffer‚ precision plus protein standard‚desalted eluate page‚ eluate page‚ wash page‚ flowthrough page‚ soluble page‚ insoluble page‚ inducted page‚ and uninducted
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8.3 POLYMORPHISMS DETECTED BY PCR Without a doubt‚ the polymerase chain reaction (PCR) represents the single most important technique in the field of molecular biology today. What PCR accomplishes in technical terms can be described very simply — it allows the rapid and unlimited amplification of specific nucleic acid sequences that may be present at very low concentrations in very complex mixtures. Within less than a decade after its initial development‚ it has become a critical tool for all practicing
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Lab Report Part II Purpose: To be familiarized with the science and techniques used to identify different types of bacteria based on their DNA sequences. Background Information: The process begins with preparing a sample. Successful identification starts with using a sample that is considered to be good. The first step is to pick up a single colony and drop it into a microcentrifuge tube. A buffer is used to dissolve the cell wall in order to extract bacterial DNA. This step may take several hours
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first one was performed to determine whose fingerprint was found at a crime scene. For this experiment‚ four-10 mL DNA samples were placed in a well of agarose gel‚ using a micropipette; the sample was also covered in a TBE buffer solution. The four samples were then put in the electrophoresis apparatus to be run through electrophoresis. After 20 minutes‚ the samples were ready and the results were documented; it was found that suspect “B” was the culprit. The second experiment was performed
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Background on Genomic DNA Isolation and Purification Generally‚ all methods involve the disruption and lysis of cells. This is followed sometimes by the removal of RNA (by RNAses‚ salt or other methods). Choosing which method to use will depend on many selection factors including: DNA is isolated from proteins by several methods including digestion of proteins by the enzyme proteinase K. Proteins are removed subsequently by salting-out‚ organic extraction‚ or binding of the DNA to a solid-phase
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III.MATERIALS AND METHODS 3.1. Collection and confirmation of Vibrio parahaemolyticus isolate Vibrio parahaemolyticus isolate was obtained from Cochin University of Science and Technology (Dr. I.S. Bright Singh) and grown in 1.5% TSB with 2% NaCl. A loop of the enriched culture was streaked onto thiosulphate-citrate-bile salt sucrose agar used for the selective isolation of vibrio strains (TCBS agar ).After 18-24hr incubation at 370c the cultures giving pure green colonies were randomly selected
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