Biochem 1 lab report

Biochem 1 lab report - Wesley Smith Monday Team 1...

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Wesley Smith Monday Team 1 Biochemistry 1 Lab Report December 7, 2009 Lab 1 (9/14/09): During this experiment, agarose gel electrophoresis was introduced as well as the introduction of automatic pipettes. Agarose gel is a quick/express way of DNA analysis which separates DNA fragments by size. Analysis of the gel gave estimate of the size of the DNA fragments. The solid agarose gel (1% in 1XTAE Buffer) was microwaved to be liquefied and approximately 25 mL was poured into the receiving tray after being mixed with 2.5 microliters of ethidium bromide. After the liquid gel was poured, approximately 20 minutes were allowed for solidification of the gel. The 20 minute waiting time was utilized for practice with automatic pipettes and they were determined to have an average of 2% error. Once the gel hardened, 5μL of ethidium bromide was added to each chamber and the tray was filled with electrode buffer. The negative electrode was situated to be on the back side of the wells in the gel since DNA has an intrinsic negative charge and passed current through the gel (approximately 120V) for about 20 minutes. The gel was analyzed using an ultraviolet light source and there were several bands present. Why were there so many bands? Because different variations of coiling present different bands; if a nuclease had been used it would have reduced the number of bands greatly to a single band. Group was successful. Lab 2 (9/21/09): In this lab, DNA was analyzed using an enzymatic digest and DNA graphing software (Delta Graph, Prizm, and Cricket Graph) was introduced. The restriction enzymes BamH1 and HindIII were used. A restriction enzyme is a site specific nuclease. Two plasmid samples were given to be analyzed, Samples A and B. Sample A was pBest luc TM and sample B was pQE31S1. Sample A had two restriction sites (HindIII and BamH1) and Sample B had three sites (1 HindIII and 2 BamH1 sites). Six samples were made to be analyzed using electrophoresis. Sample 1 was a control consisting of 1μL NEB2 X 10 reaction buffer, 5μL plasmid sample A, and 4μL of purified water. Sample 2 had the addition of 1μL of BamH1 and Sample 3 had the addition of 1μL HindIII. Sample 3 was also a control using instead 5μL of plasmid sample B,
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Biochem 1 lab report - Wesley Smith Monday Team 1...

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