Long PCR stuck in the wells :-( (fwd)

Robert Preston rapr at MED.PITT.EDU
Sat May 28 13:43:17 EST 1994


Forwarded message:
> From: hdang at cns.neusc.bcm.tmc.edu (Hong Dang)
> Subject: Long PCR stuck in the wells :-(
> Date: 27 May 1994 21:42:08 GMT
> Dear Bionetters:
> We are trying inverse PCR of 6 kb plasmid according to Barnes' mixing polymerases
> strategy. Some primer sets worked real well, while others not so well. However, 1
> set of primers produced either nothing or heavy EtBr band stuck in the well.
> 
> Anybody has any clue as what the stuff is and how to "move" it down to the right
> size PCR band?
> 
> Any input is greatly appreciated.
> 
> Hong
> hdang at channel.neusc.bcm.tmc.edu
> 
Hong, Barnes himself posted (search the archives) that too many cycles
makes for junk in the gel wells: he said to cut it to 16 cycles to avoid
that.  I have no idea why, myself.

Rob Preston
rapr at med.pitt.edu




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