Given the following DNA sequence for a gene ispD from Yersinia pestis, and the s

Question

Given the following DNA sequence for a gene ispD from Yersinia pestis, and the sequences of the forward primer A and the reverse primer A, what size band (in base pairs) would you expect to see on a DNA gel after you conducted PCR with this primer set? What size band would you expect to see with forward primer B and reverse primer B? (HINT: If you’re struggling, take a look at https://www.youtube.com/watch?v=tUyBwiyMsSU, at least the first 11 minutes or so, for help. A forward primer matches the target sequence on the forward strand, while the reverse primer is the reverse complement of the target sequence on the forward strand. )

Primer Set A: Forward: 5’ GGTGGATTGCCCTAAGCAGT 3’

                        Reverse: 5’ TCTGGGCGCGTCACTTTAAT 3’

Primer Set B: Forward: 5’ TGGTGGATTGCCCTAAGCAG 3’

                        Reverse: 5’ ACCCCTTCTCTTAACGCACG 3’

Yersinia pestis ispD sequence:

5’ atgagtaacttcgcagtttcccttcctgaagtgatcgctgtattaccggctgcgggtattggtagccgtatgttggtggattgccctaagcagtatttaactgtggggggcaaaacaatcattgaacatgctattttttctttgcttcaccacccacgaattcagcgggttatcgttgtgatccatccgcaggacacacaattctctaggttgtccgttgcgcaggatccacgtatcagtacagtttacggtggcgatcaacgggctaactccgtgatggcgggtttacaattggcagggcaggctgaatgggtgttagttcatgatgcggcacgcccctgtttgcaccttgatgatctcagccggctgttatcgattaccgaatgcagtcaggtggggggaattctggcggcccctgtgcgtgatacgatgaaacgtgccgagccgggtattcaagccatcgctcatacggtggatcgtcaggacctgtggcatgcgctgacgcctcaacttttcccgctagaattattaaaattgtgcttatcccgtgcgttaagagaaggggtggcggtgactgatgaggcctctgcattagagcattgcggttatcatccgatattggttaccggccgttctgataatattaaagtgacgcgcccagaagatctggcattggcggagttttatttaacccagcggcagtctctcaataacgacagtctctga

                                  3’

Explanation / Answer

Primer A

Forward primer 5’ GGTGGATTGCCCTAAGCAGT 3’

Complementary strand for Reverse primer 5’ TCTGGGCGCGTCACTTTAAT 3’ 5' attaaagtgacgcgcccaga 3'

5’ atgagtaacttcgcagtttcccttcctgaagtgatcgctgtattaccggctgcgggtattggtagccgtatgttggtggattgccctaagcagtatttaactgtggggggcaaaacaatcattgaacatgctattttttctttgcttcaccacccacgaattcagcgggttatcgttgtgatccatccgcaggacacacaattctctaggttgtccgttgcgcaggatccacgtatcagtacagtttacggtggcgatcaacgggctaactccgtgatggcgggtttacaattggcagggcaggctgaatgggtgttagttcatgatgcggcacgcccctgtttgcaccttgatgatctcagccggctgttatcgattaccgaatgcagtcaggtggggggaattctggcggcccctgtgcgtgatacgatgaaacgtgccgagccgggtattcaagccatcgctcatacggtggatcgtcaggacctgtggcatgcgctgacgcctcaacttttcccgctagaattattaaaattgtgcttatcccgtgcgttaagagaaggggtggcggtgactgatgaggcctctgcattagagcattgcggttatcatccgatattggttaccggccgttctgataatattaaagtgacgcgcccagaagatctggcattggcggagttttatttaacccagcggcagtctctcaataacgacagtctctga 3’

Size of the sequence formed - 588 bp

Primer B

Forward primer 5’ TGGTGGATTGCCCTAAGCAG 3’

Complementary strand for Reverse primer 5’ACCCCTTCTCTTAACGCACG 3’ 5' cgtgcgttaagagaaggggt 3'

5’ atgagtaacttcgcagtttcccttcctgaagtgatcgctgtattaccggctgcgggtattggtagccgtatgttggtggattgccctaagcagtatttaactgtggggggcaaaacaatcattgaacatgctattttttctttgcttcaccacccacgaattcagcgggttatcgttgtgatccatccgcaggacacacaattctctaggttgtccgttgcgcaggatccacgtatcagtacagtttacggtggcgatcaacgggctaactccgtgatggcgggtttacaattggcagggcaggctgaatgggtgttagttcatgatgcggcacgcccctgtttgcaccttgatgatctcagccggctgttatcgattaccgaatgcagtcaggtggggggaattctggcggcccctgtgcgtgatacgatgaaacgtgccgagccgggtattcaagccatcgctcatacggtggatcgtcaggacctgtggcatgcgctgacgcctcaacttttcccgctagaattattaaaattgtgcttatcccgtgcgttaagagaaggggtggcggtgactgatgaggcctctgcattagagcattgcggttatcatccgatattggttaccggccgttctgataatattaaagtgacgcgcccagaagatctggcattggcggagttttatttaacccagcggcagtctctcaataacgacagtctctga 3’

Size of the sequence formed - 493 bp

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