Posted 07 July 2011 - 07:43 AM
Just try once: use normal Taq, do the PCR again, and try ligate again into your vector. If this succeed, this means you might did sthg wrong during the A-tailing procedure. If not, it could be your vector/ligation problem.
Just 2 cents.
Expecting the world to treat you fairly because you are a good person is like expecting the lion not to attack you because you are a vegetarian.
..."best of our knowledge, as far as we know this had never been reported before, though I can't possible read all the published journals on earth, but by perform thorough search in google, the keywords did not match any documents"...
"what doesn't kill you, makes you stronger"---Goddess Casandra reminds me to be strong
"It's all just DNA. Do it."---phage434