Home FAQs Can the cloning kit be used with inserts containing 5´ or 3´ overhangs greater than the single-base overhang achieved by PCR with Taq DNA polymerase?

FAQ: Can the cloning kit be used with inserts containing 5´ or 3´ overhangs greater than the single-base overhang achieved by PCR with Taq DNA polymerase?

Yes, but with lower efficiencies. The polishing components are optimized for removal of a single base overhang. As an extreme test example, cloning a DNA fragment possessing a 4-base, 5´ overhang requiring a fill in and a 5-base, 3´ overhang requiring a chew back resulted in 10-fold fewer transformants than blunt end or single-base overhang DNA fragments.