Pfu Polymerase - Should die a slooowww death (May/20/2007 )
Am I the only one who hates this stupid polymerase? 
Perhaps I have been spoiled in the past but the new lab I work, the only polymerase we have with proofreading activity is Pfu. And it is sooooooo slow and the yields of DNA are lower than I am used to. I can do my work but it so annoying doing PCRs that should take 2hrs taking 5+ hours. My new project I will be amplifying large pieces of approximately 6-10kb of genomic DNA. It will be very time sensitive to get all the cloning done. If I have to use Pfu I will seriously go nuts.
-the_pwnererr-
QUOTE (the_pwnererr @ May 20 2007, 04:22 PM)
Am I the only one who hates this stupid polymerase? Perhaps I have been spoiled in the past but the new lab I work, the only polymerase we have with proofreading activity is Pfu. And it is sooooooo slow and the yields of DNA are lower than I am used to. I can do my work but it so annoying doing PCRs that should take 2hrs taking 5+ hours. My new project I will be amplifying large pieces of approximately 6-10kb of genomic DNA. It will be very time sensitive to get all the cloning done. If I have to use Pfu I will seriously go nuts.
Perhaps I have been spoiled in the past but the new lab I work, the only polymerase we have with proofreading activity is Pfu. And it is sooooooo slow and the yields of DNA are lower than I am used to. I can do my work but it so annoying doing PCRs that should take 2hrs taking 5+ hours. My new project I will be amplifying large pieces of approximately 6-10kb of genomic DNA. It will be very time sensitive to get all the cloning done. If I have to use Pfu I will seriously go nuts.which company's pfu are you using?
-T. reesei-
QUOTE (T. reesei @ May 20 2007, 05:40 PM)
QUOTE (the_pwnererr @ May 20 2007, 04:22 PM)
Am I the only one who hates this stupid polymerase? Perhaps I have been spoiled in the past but the new lab I work, the only polymerase we have with proofreading activity is Pfu. And it is sooooooo slow and the yields of DNA are lower than I am used to. I can do my work but it so annoying doing PCRs that should take 2hrs taking 5+ hours. My new project I will be amplifying large pieces of approximately 6-10kb of genomic DNA. It will be very time sensitive to get all the cloning done. If I have to use Pfu I will seriously go nuts.
Perhaps I have been spoiled in the past but the new lab I work, the only polymerase we have with proofreading activity is Pfu. And it is sooooooo slow and the yields of DNA are lower than I am used to. I can do my work but it so annoying doing PCRs that should take 2hrs taking 5+ hours. My new project I will be amplifying large pieces of approximately 6-10kb of genomic DNA. It will be very time sensitive to get all the cloning done. If I have to use Pfu I will seriously go nuts.which company's pfu are you using?
Fermentas. I know some are better than others. Of all the proofreading polymerases I've used, I liked Phusion the best. Its the best polymerase money can buy IMO. But there are also other good ones out there.
-the_pwnererr-
Why not buy a different polymerase then? Or do your PCR overnight?
-vairus-
QUOTE (vairus @ May 20 2007, 11:40 PM)
Why not buy a different polymerase then? Or do your PCR overnight?
change your polymerase or choose overnight
-T. reesei-