The underlined regions in the following sequence indicate the annealing sites of
ID: 91629 • Letter: T
Question
The underlined regions in the following sequence indicate the annealing sites of two primers to be used in a polymerase chain reaction. 5’CCGGTGCTGCAATGCTTTTTAGACGGCTTGCTGTCCGCCTTATCCACATGACTCTCGGCA ACGGATATCTCGGCTCTCGCATCGATGAAGAACGTAGCGAAATGCGATACCTGGTGTGAAT TGCAGAATCCCGCGAACCATCGAGTCTTTGAACGCAAGTTGCGCCCGAGGCCATTCGGCC GAGGGCACGCCTGCCTGGGCGTCACGCAAAACACGCTCCCAACCCCCCTCATCGGGGAG CGGGATGCGGCATGTGGCTTTCCGTCACGCAAGTGGCGGTGGGCCGAAGATATGGCTGC CGGCGTGGTGGGCGTCATCGCTTTGTATCGTGCCGGACACAGCG 3’
Which of the following primer pairs should be used to amplify this sequence of DNA? Explain your answer. (2 points)
A. 5’ CCGGTGCTGCAATGCTTTTTA 3’ and 5’ TGTATCGTGCCGGACACAGCG 3’
B. 5’ CCGGTGCTGCAATGCTTTTTA 3’ and 5’GCGACACAGGCCGTGCTATGT 3’
C. 5’ ATTTTTCGTAACGTCGTGGCC 3’ and 5’ TGTATCGTGCCGGACACAGCG 3’
D. 5’ CGCTGTGTCCGGCACGATACA 3’and 5’ CCGGTGCTGCAATGCTTTTTA 3’
Explanation / Answer
Ans- Option (D)
reverse primer- 5’ CGCTGTGTCCGGCACGATACA 3’
forward primer-5’ CCGGTGCTGCAATGCTTTTTA 3’
5’CCGGTGCTGCAATGCTTTTTAGACGGCTTGCTGTCCGCCTTATCCACATGACTCTCGGCA
5’CCGGTGCTGCAATGCTTTTTA3’-----
ACGGATATCTCGGCTCTCGCATCGATGAAGAACGTAGCGAAATGCGATACCTGGTGTGAAT TGCAGAATCCCGCGAACCATCGAGTCTTTGAACGCAAGTTGCGCCCGAGGCCATTCGGCC GAGGGCACGCCTGCCTGGGCGTCACGCAAAACACGCTCCCAACCCCCCTCATCGGGGAG CGGGATGCGGCATGTGGCTTTCCGTCACGCAAGTGGCGGTGGGCCGAAGATATGGCTGC CGGCGTGGTGGGCGTCATCGCTTTGTATCGTGCCGGACACAGCG 3’
--------------- '3 ACATAGCACGGCCTGTGTCGC 5' (Reverse primer)
DNA replicates from 5' to 3' direction. Forward primer has same sequence present at 5' end of the DNA called sense primer and it will amplify top strand. But reverse primer is complementary to the sequence present at 3' terminal of the DNA which will amplify bottom strand ( reverse primer has similar sequence present at 5' terminal of bottom strand)
5'------------3'
3'-----------5'
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