Lab I Report after peer review

Lab I Report after peer review - Lauren Allen February 12,...

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Lauren Allen February 12, 2007 Dr. Tom Seiflein Molecular Biology Lab I & II
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Determination of a Bacterial Plasmid via Phenotypic and Genotypic Tests RESULTS Table 1: Expected and Observed Phenotypes of E. Coli with Plasmids Containing Spliced DNA on Agars Pre-treated with Antibiotics (GROW or GLOW?) Plus means growth, minus means no growth, Yes means the culture glowed under UV light, and No means that it did not. Table 2: Fragment Length and Relative Mobility After Electrophoresis in Both Lambda DNA and an Unknown Cut DNA Sample, Both Cut With the HIN DIII Enzyme Table 3: Expected Fragment Lengths of the Plasmid DNA Cuts by Restriction Enzymes Compared to the Unknown Sample DESCRIPTION OF DATA Bacteria that contain the pAMP plasmid did not glow on any of the agars, and grew on all agars except the agar containing the kanamyacin antibiotic. Bacteria containing the pKAN plasmid did not glow on any of the agars, and grew on all of the agars except for those with the ampicillin and the ampicillin/arabinose antibiotic on them. Bacteria containing the pGLO plasmid grew on all agars except the agar with the kanamyacin and did not glow on any but the ampicillin/arabinose containing agars. The bacteria with the unknown cut plasmid DNA that was tested grew on all the agars except the kanamyacin containing agar and glowed on the ampicillin/arabinose agar plates. This is shown in Table 1. For the genotypic test an alkaline lysis mini-prep was required (since we were only isolating a small amount of plasmid). This sample of plasmid was then dyed and placed in electrophoresis agarose gel wells. Another sample of this plasmid was cut by another restriction enzyme, as well as a sample of Lambda DNA. These were both put in
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the wells. A sample of uncut Lambda DNA was as well. Electrophoresis was then allowed to occur, and the relative mobility of the fragments (in cm) was then compared between the cut Lambda DNA (of which the base pair lengths are very well known and standard) and the cut unknown plasmid. This was to help determine the base pair length of the cut plasmid DNA. The lambda fragments traveled 2.2 cm, 2.5 cm, 2.9 cm, 3.4 cm, 4.1 cm, 4.3 cm, and 6.4 cm. Their base pair lengths, respectively, are 23130bp, 9416bp, 6557bp, 4361bp, 2322bp, 2027bp, and 564bp. The unknown sample traveled 3.1cm,
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This note was uploaded on 04/14/2008 for the course BIOSCI 0060 taught by Professor Ruthermund during the Spring '08 term at Pittsburgh.

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Lab I Report after peer review - Lauren Allen February 12,...

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