Partner’s Name: Manan
Assignment 2
POLYMERASE CHAIN REACTION
Total Marks: 12
1. (2 marks) In this experiment, you have amplified the D1S80 locus by PCR. Explain the advantages of using this locus to distinguish one person from another. Do you think you could use a coding gene for the same purpose? Clearly explain your answers.
D1S80 locus is placed on the short arm of the chromosome 1. This locus does not code for the arrangement for protein, yet it codes for a series of tandem repeats of 16 bp in human. Distinctive number of this allele has different number of repeats. These quantities of repeats are exceptional to every human. Primer is bind to this repeats and after amplification this makes fragments of different sized which can be seen on the agarose gel. From the extent of the fragment one can figure the number of repeats by following equation.
113 bp + (n)(16 bp) + 32 bp = fragment size
Where, n = number of repeats.
I think coding gene cannot be utilized as coding genes code a sequence of protein. People have a few quantities of same proteins which implies that every human has a few same coding sequences. If we use coding gene for this purpose, it makes impossible for us to differentiate humans.
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2. (8 marks) Analyze your results. Your analysis must include:
- the picture of the gel; clearly label the lanes and show all your measurements
- a table summarizing your TECDNA data
- a graph of the TECDNA data; clearly show how you use this standard curve to calculate the size of your PCR product(s); include one example of each of your calculations;
- a final statement, including the number of repeats present in your D1S80 alleles.
Observation Table 1: The distance traveled by each fragments of lambda DNA on basis of molecular weight.
The last 5 fragments of 1kb gene ruler from smallest to largest
Molecular weight of 1 kb gene ruler (bp)
Log