Below is the nucleotide sequence for a template strand of DNA and the sequences

Question

Below is the nucleotide sequence for a template strand of DNA and the sequences for the forward and reverse primers. Template forward strand: 5'-ATCGGCTCGCTATCATTGCATCAACAGGCATTCAACAATGCTGCTCAGCTCA -3 Template reverse strand: 3'- TAGCCGAGCGATAGTAACGTAGTTGTCCGTAAGTTCTTACGACGAGTCGAGT-5' Forward primer: 5'- CGCTATC -3' Reverse primer: 3'- GACGAGT -5' What is the DNA sequence for the DNA that will be produced from PCR using this template DNA and primers? Remember that the primer becomes part of the PCR product. 3'- GCGATAGGACGAGT -5 5'-ATTGCATCAACAGGCATTCAACAATG -3 3'- TAACGTAGTTGTCCGTAAGTTCTTAC-5 5'- CGCTATCATTGCATCAACAGGCATTCAACAATGCTGCTCA-3 5'- CGCTATCATTGCATCAACAGGCATTCAACAATGCTGCTCAGCTCA -3 3-TAGCCGAGCGATAGTAACGTAGTTGTCCGTAAGTTCTTACGACGAGT -5

Explanation / Answer

Answer: The Option 4 is the correct Answer.

Explanation:

The Double Stranded Template DNA splits into two single strands as follows:

5' - ATCGGCTCGCTATCATTGCATCAACAGGCATTCAACAATGCTGCTCAGCTCA - 3'

and

3' - TAGCCGAGCGATAGTAACGTAGTTGTCCGTAAGTTCTTACGACGAGTCGAGT - 5'

After this the primers anneal.

The forward primer 5'-CGCTATC-3' specifically anneals at this complimentary region of the template reverse strand as shown below:

                  5'-CGCTATC-3'                                                                                                            3' - TAGCCGAGCGATAGTAACGTAGTTGTCCGTAAGTTCTTACGACGAGTCGAGT - 5'

And the reverse primer 3' - GACGAGT - 5' specifically anneals to the complimentary region of the forward template strand as shown below:

5' - ATCGGCTCGCTATCATTGCATCAACAGGCATTCAACAATGCTGCTCAGCTCA - 3'

                                                                                     3' - GACGAGT - 5'

After annealing as shown above, the strands are extended from the forward primer and reverse primers.

It is important to remember that A & T and G & C are the complimentary pairs.

After certain steps the shortening of the strands take place and result in the final DNA sequence as given below - Option No. 4 in the question.

5' - CGCTATCATTGCATCAACAGGCATTCAACAATGCTGCTCA - 3'

3' - GCGATAGTAACGTAGTTGTCCGTAAGTTCTTACGACGAGT - 5'

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