request follow-up on SmaI (PCR subcloning woes cont.)

T. S. Pillay tpillay at
Thu Mar 10 11:19:05 EST 1994

In article <940309102207.54436 at> ,
txpljfg at UABCVSR.CVSR.UAB.EDU writes:
>I have sinced stopped using blunt end cloning and I now use TA cloning
>for all my PCR products.  I incubate blunt-ended pUC19 with 2mM dTTP in
>standard 1X PCR buffer for three hours at 75 degrees, ppt with ammonium
>acetate and isopropanol, wash a couple of times and ligate normally.

Could you please let us have your detailed  protocol for making pUC19
with T-overhangs?
Many thanks

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