25 February 2023 3 8K Report

I want to create a recombinant plasmid with the insert of size 1128 bp and the plasmid pET-15b of size 5708 bp. DNA insert and plasmid vector was double digested with BamH1 and BIpI restriction enzymes. I have done ligation reactions in 3:1, 5:1 and 7:1 DNA insert:vector ratios. Attached is the agarose gel image with the following description:

LANE 1 -1 kb DNA ladder

LANE 3 - Double digested pET-15b plasmid

LANE 4 - Double digested DNA insert.

LANE 6 - Ligated product of 3:1 ratio

LANE 7 - Ligated product of 5:1 ratio

LANE 8 - Ligated product of 7:1 ratio

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