cloning a pfu amplified fragment in T vector
(by rahimpour_a from yahoo.com)
Sat Feb 6 06:38:18 EST 2010
I need to clone a pfu amplified fragment in t-vector, in the vector manual it is said that pfu pcr product must be purified before a tailing by tag because pfu can remove a overhangs by its proofreading activity, but in commercial high fidelity enzyme mixes which include a proofreading enzyme with tag we can use pcr products directly for t/a ligation, so why in the latter case A overhangs are not removed?
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