Question

The sequence of the fruit fly gene, cg8216, is listed below.

>cg8216

ATGAGCATTTTTCCGAGGTGGGCAGCCATGCTGCTGCTCCTGGGCCTGGC

CCACCAGCTGGACCCCTCGCTCGCGGCGTCCGCATCCTCGTTCGCATCCG GAAACGCCTGGCAGCGCGCCCTATTTGGCCGGGAATCGCGCAACTTGATG CACCGCAGAGCCCCGTTTTCCGGAGCCGCTGATCTGGGCCTTGACGAATA CCTGGGCCCCTATGGCGCCGTCGAGCAGGAGCAGCACCAGCCGCATCCTC CGCCGCAGCAGATGCGCCAACAGACCGAAGTTTACGGAATCGTGGAGCCG CTGATTGAGGACACGCCCTGTGCCGACCGACCCTGCCTCCTCAACGACGA TTGCTGTCCTTCCGGAGTTTGCGTCAGCACATACGGCGAAGGCAAATGCG TATATGTCTTTGGGCGTCAGCGAGATCTGTGCCAGGGACATGCGGACTGT CCGCAGGGCAGCTCCTGCATGCTGGTCCCCCAGGAGGGAGCGTGGCGGTG TGAGCCGAGCGTGGAATCGGGAGGATCCACGTCGCTGCTGGAGGGTATAT TCGGAGCGAAGGAGAGGCAACCGCTGGGCAGCGAGTGCAGCAGCAGCAGC GACTGCCAGGTGATCAACGGAATGTGCTGTCAACAGCAGCGTTTGCACCA CCGGGCAGCCATTAAGCTGAGCTGTGGCTACTTTCGCGATGCCTTCGACT GTGTAGATATGGTCGGAGCAGAGCATCGACGCAACTGA

We used this DNA sequence, within a plasmid, as a positive control for our PCR reactions.

Using the given primer sequences, locate the complementary location of each primer within the sequence.

Primer sequences:

cg8216 IS-1 F           TTCCGAGGTGGGCAGCCAT

cg8216 IS-1 R          CTGGGGGACCAGCATGCAG

Using this process, theoretically validate that the predicted resulting PCR product is approximately 471 base pairs. Explain your answer.

Answer 1

5'ATGAGCATTTTTCCGAGGTGGGCAGCCATGCTGCTGCTCCTGGGCCTGGC

CCACCAGCTGGACCCCTCGCTCGCGGCGTCCGCATCCTCGTTCGCATCCG GAAACGCCTGGCAGCGCGCCCTATTTGGCCGGGAATCGCGCAACTTGATG CACCGCAGAGCCCCGTTTTCCGGAGCCGCTGATCTGGGCCTTGACGAATA CCTGGGCCCCTATGGCGCCGTCGAGCAGGAGCAGCACCAGCCGCATCCTC CGCCGCAGCAGATGCGCCAACAGACCGAAGTTTACGGAATCGTGGAGCCG CTGATTGAGGACACGCCCTGTGCCGACCGACCCTGCCTCCTCAACGACGA TTGCTGTCCTTCCGGAGTTTGCGTCAGCACATACGGCGAAGGCAAATGCG TATATGTCTTTGGGCGTCAGCGAGATCTGTGCCAGGGACATGCGGACTGT CCGCAGGGCAGCTCCTGCATGCTGGTCCCCCAGGAGGGAGCGTGGCGGTG TGAGCCGAGCGTGGAATCGGGAGGATCCACGTCGCTGCTGGAGGGTATAT TCGGAGCGAAGGAGAGGCAACCGCTGGGCAGCGAGTGCAGCAGCAGCAGC GACTGCCAGGTGATCAACGGAATGTGCTGTCAACAGCAGCGTTTGCACCA CCGGGCAGCCATTAAGCTGAGCTGTGGCTACTTTCGCGATGCCTTCGACT GTGTAGATATGGTCGGAGCAGAGCATCGACGCAACTGA 3'

In the above template i have m entioned the complementary primer binding sites in bold and italuc letters for forward primer at 5' end and reverse primer at 3' end

Predicted PCR product size would be approximately 471 bp and it can be identified by counting the number of bases that are prsent at the start site of forward primer and end nucleotide of reverse primer