did you got some amplification ? How did you do that?I did a PCR once with primers one at 46 and the other one at 71 (long story, but I couldn't design them better no matter what I did, and I struggled quite a lot). Worked perfectly....but I'm the one with a too complicated PCR program
Andreea
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18 replies to this topic
#16
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Posted 26 August 2012 - 08:09 AM
#17
ascacioc
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Posted 26 August 2012 - 03:49 PM
see my previous posts, after I said that I did that I also posted a protocol I use for Phusion PCR.
Andreea
Andreea
#18
Trof
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Posted 04 October 2012 - 03:00 AM
Open source is a nice thing and surely if someones digs out the right composition by trying and googling good for him, on the other hand I have no problem with paying company for a good product they developed 
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#19
brassmonkey
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Posted 03 March 2014 - 02:31 PM
I know this isn't at all timely, but Andreea mentioned SB buffer. The same authors (Brody and Kern) later published on LB buffer (Lithium instead of sodium). Now I run my gels for 10 min at 300V in Owl chambers rated to 150V and everything is great. LB definitely out-performs SB. Also a phusion lover, I'm going to try your touchdown protocol. I've become less-enamored with touchdowns over the last couple of years, but I'll entertain your protocol.
