Give the sequence of a reverse primer used to insert a BamHI site (GGATCC) at the 3' end of the following sequence:

Question

Give the sequence of a reverse primer used to insert a BamHI site (GGATCC) at the 3' end of the following sequence:

5' CTGAACGTCCAAGGCTTAA 3'
3' GACTTGCAGGTTCCGAATT 5'

Make sure you indicate your primers polarity.

Answer 1

To insert a BamHI site (GGATCC) at the 3' end of the given sequence, we need to design a reverse primer that is complementary to the original sequence and includes the BamHI site.

Here are the steps to design the reverse primer:

1. Write down the original sequence:
5' CTGAACGTCCAAGGCTTAA 3'

2. Find the reverse complement of the sequence by pairing each base with its complementary base:
5' CTGAACGTCCAAGGCTTAA 3'
---> GACTTGCAGGTTCCGAATT 3'

3. Add the BamHI site (GGATCC) to the 3' end of the reverse complement sequence:
GACTTGCAGGTTCCGAATT GGATCC

4. Reverse the sequence to match the reverse primer polarity:
CCATGTTCGGAACTTAAGGATCC

Therefore, the reverse primer sequence, with the BamHI site inserted at the 3' end, is:
5' CCATGTTCGGAACTTAAGGATCC 3'

Make sure to indicate the polarity of the primer when ordering or using it in experiments. In this case, the reverse primer polarity is 5' to 3'.