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PCR after T4 ligation?


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#1 jesscalleros

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Posted 30 July 2014 - 03:15 PM

Greetings friends and colleagues, I'm just wondering, if It's possible to make a pcr after a t4 ligation, without preivious EtOH precipitation? I mean... The T4 buffer could be a problem for the PCR reaction?

Thanks for read!

#2 phage434

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Posted 30 July 2014 - 03:41 PM

You can use vanishingly small amounts of your ligation product in your PCR reaction, so the effect will be near zero.



#3 jesscalleros

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Posted 30 July 2014 - 03:49 PM

Well Amaizing! That's sounds obvious, but I had to be sure, thanks a lot!

#4 Bio-Lad

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Posted 10 August 2014 - 07:32 PM

I've done this before.  I used 1uL of my ligation in a 50uL PCR reaction (using NEB's Phusion polymerase and HF buffer) and it worked great for me.


Try pLOT, the a free plasmid mapping tool!

http://plasmidplotter.blogspot.com/





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