2. A. You want to use PCR to amplify the entire DNA shown in the figure below. O
ID: 217369 • Letter: 2
Question
2. A. You want to use PCR to amplify the entire DNA shown in the figure below. Of the listed primers, choose the pair that will allow you to amplify this stretch of DNA by PCR. (9 points total) DNA to be amplified: 5' -CGATTCGAGCCATTCGAACTCGATCAGCCGATTCGATCAACCTTGGACAGTCAG-3 3'-GCTAAGCTCGGTAAGCTTGAGCTAGTCGGCTAAGCTAGTTGGAACCTGTCAGTC-5 , Primers: (1) 5'-GCTAAGCTCGGTA-3, (2) 5'-GAACCTGTCAGTC-3' (3) 5'-CTGACTGTCCAAG-3 (4) 5'-GACTGACAGGTTC-3' (5) 5'-CTTGGACAGTCAG-3' (6) 5'-cGATTCGAGCCAT-3' (7) 5'-ATGGCTCGAATCG-3 (8) 5'-TACCGAGCTTAGC-3' Answer: I will need primer numberand primer number3 points each) B. How many PCR cycles do you need to perform in order to obtain 128 double-stranded copies of a DNA molecule, if you start with a single molecule of double-stranded DNA and assume that your PCR steps are 100% efficient? (3 points)Explanation / Answer
Q.2 (A)
Sense strand runs from 5' to 3' direction and is complementary to antisense strand which runs from 3' to 5' direction.
Forward primer anneal to the anti-sense strand . And hence we can choose No.6 primer for forward reaction.
Reverse primer anneal to the sense strand. And hence we choose No.3 primer for reverse reaction.
Q.2 (B)
After one cycle PCR, 2 dsDNA is obtained from single double stranded DNA.
27 = 128 dsDNA.
7 PCR Cycle is required to obtain 128 dsDNA.
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