[Q]help-TA cloning after PCR

601 h971409d at eds.ecip.nagoya-u.ac.jp
Wed Jan 7 14:08:56 EST 1998


Hello, eveyone.
I want to know about how to do TA cloning well.  I have just made
a T vector by the following protocol.

pBluscript 10micrograms -(EcoRV digestion) - (Et-OH precipitation)- 
-(70degrees centigrade, 2hrs with dTTP in thrmocycler)-purification

But my vectors didn't work well.  The sample before ligation (no insert)
could make a lot of white colonies.  Maybe to add T residue to pBluscript
was not enough, I think.

Please give me som or ideas advice. Thank you in advance.
Thank you.

-- 
Shin Yamamoto
h971409d at eds.ecip.nagoya-u.ac.jp



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