[Q]help-TA cloning after PCR
verkhovs at penger.helsinki.fi
Fri Jan 23 10:35:47 EST 1998
I have tried In vitrogen TA cloning kit with a very good luck. Of course, it
is not very nice to depend only on a kit and they give an open vector so you
can not use it to transform cells and have your own suply but works very,very
University of Helsinki
email. barquera at rock.helsinki.fi
> 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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