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Forum: 454 Pyrosequencing 09-04-2011, 11:42 PM
Replies: 69
Views: 27,942
Posted By Cambridge454
It turned out that when the AMPure XP beads near...

It turned out that when the AMPure XP beads near their expiration date we were seeing this problem. Our library sizes returned to normal when we switched to a new bottle of beads.
Forum: 454 Pyrosequencing 03-18-2010, 02:14 AM
Replies: 69
Views: 27,942
Posted By Cambridge454
I am checking my library using the Bioanalyzer...

I am checking my library using the Bioanalyzer High Sensitivity DNA chip as recommended in the protocol. Do you think the fact that for the Rapid Libraries you are starting with 500ng instead of 5ug...
Forum: 454 Pyrosequencing 03-16-2010, 06:23 AM
Replies: 69
Views: 27,942
Posted By Cambridge454
Rapid Library length problems

I am also having problems with the Rapid Library producing really long average fragment lengths >900bp. I use the recommended nebulization settings, which worked perfectly well for old version...
Forum: 454 Pyrosequencing 11-09-2009, 06:37 AM
Replies: 9
Views: 5,052
Posted By Cambridge454
Thanks for the advice. I think this might be my...

Thanks for the advice. I think this might be my problem, as I usually pour off the isopropanol supernatant, add the 1x Enhancing Fluid TW, vortex, vortex, vortex, break clumps with glass rod, vortex,...
Forum: 454 Pyrosequencing 11-08-2009, 11:28 PM
Replies: 9
Views: 5,052
Posted By Cambridge454
Hi Jason, Yes, I have read and followed the...

Hi Jason,

Yes, I have read and followed the instructions regarding the additive in the Technical Bulletin 011-2009. There appears to be no precipitate in the Live Amp Mix, it only reappears again...
Forum: 454 Pyrosequencing 11-06-2009, 06:42 AM
Replies: 9
Views: 5,052
Posted By Cambridge454
emPCR additive

I have not once been able to fully dissolve the precipitate in the emPCR additive, even after heating to 55C. It does not seem to affect the sequencing results but I have noticed especially with the...
Forum: Introductions 11-06-2009, 06:25 AM
Replies: 373
Views: 169,088
Posted By Cambridge454
Hi everyone, My name is Shilo and I work in...

Hi everyone,

My name is Shilo and I work in the Sequencing Facility at the University of Cambridge. We got our 454 in April 2008 and it sure was a big adjustment after many, many years of Sanger...
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