Mabel Akinwumiju

Molecular Cell Biology Lab

Due: December 4, 2007

TA: Xiaoli Zhang

Group #6

Polymerase Chain Reaction

Materials and Methods:

In order to perform polymerase chain reaction, the following cocktail was made:  DNA template, deoxynucleotide triphosphates (dNTP), Taq polymerase, two primers, and a buffer solution.  Before starting the PCR, first four samples were collected from the prime suspects and 19 uL “cocktail” was added.  1 uL of DNA from each suspect was placed into separate tubes, then 89 uL dH20, 6 uL dNTP, 3ul of forward primer, 3 uL of reverse primer, 1 uL Taq polymerase, and 12 uL 10X PCR buffer was added into each eppendorf tube.  After thoroughly mixing, the samples were placed into the PCR tubes and then placed into the thermocyler.

  After the PCR cycle was preformed 2 uL of tracking dye was added to each sample and then they each were loaded into 7% acrylamide gel in 1X TBE solution for gel electrophoresis.  The gel was run for approximately 1 ½  hours at 240 volts, and then the gel was placed in an ethidium bromide solution for 10 minutes to help stain the DNA.  Then the TA photographed it under a UV light.

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Results:

After the gel electrophoresis procedure was complete, we were able to generate the PCR result

The figure on the picture found in the appendix depicts the results that were obtained from the gel electrophoresis done for the PCR. The lanes were numbered from left to right as Cs (crime scene), 4, 3, 2, and 1.

        The banding patterns were clear for all five lanes. The lane that contained the DNA from the crime scene had a total of three bands near the well of the gel one band at 16 mm (millimeter), one at 18 mm, ...

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