[Q]help-TA cloning after PCR

Jiqing Peng peng.36 at postbox.acs.ohio-state.edu
Thu Jan 8 20:48:10 EST 1998


In article <h971409d-0801980412270001 at news.cc.nagoya-u.ac.jp> h971409d at eds.ecip.nagoya-u.ac.jp (601) writes:
In article <h971409d-0801980412270001 at news.cc.nagoya-u.ac.jp> h971409d at eds.ecip.nagoya-u.ac.jp (601) writes:
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>From: h971409d at eds.ecip.nagoya-u.ac.jp (601)
>Newsgroups: bionet.molbio.methds-reagnts
>Subject: [Q]help-TA cloning after PCR
>Date: 7 Jan 1998 19:08:56 GMT
>Organization: Nagoya University
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>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.


Why don't you buy a TA cloning kit.  It works perfect.




>Thank you.

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




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