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Forum: 454 Pyrosequencing 12-14-2009, 08:00 AM
Replies: 0
Views: 2,172
Posted By boss_hoss
New 454 emulsion creation

Hey all-
Has anyone tried using the IKA UltraTurrax to create 454 emulsions? The 454 Junior press release made it seem the protocol has shifted towards this method.

"...instead of a tissue lyser...
Forum: 454 Pyrosequencing 10-09-2009, 10:53 AM
Replies: 21
Views: 11,737
Posted By boss_hoss
we had a similar problem with specifically SVE...

we had a similar problem with specifically SVE setups, where we were seeing a ton of broken wells and poor enrichments. it turned out to not only be an oil lot problem, but an oil problem that...
Forum: 454 Pyrosequencing 10-08-2009, 12:46 PM
Replies: 21
Views: 11,737
Posted By boss_hoss
have you seen any broken emulsion wells? do your...

have you seen any broken emulsion wells? do your library traces look normal? what is your enrichment percentage specification range?
Forum: 454 Pyrosequencing 06-24-2009, 07:39 AM
Replies: 3
Views: 3,030
Posted By boss_hoss
emPCR additive

Anybody have an idea what the emPCR additive actually is?

DMSO? Betaine? Tw20? Magic?
Forum: 454 Pyrosequencing 06-24-2009, 07:36 AM
Replies: 6
Views: 2,868
Posted By boss_hoss
I agree with genomeseeker - if the raw wells are...

I agree with genomeseeker - if the raw wells are low, this indicates a problem with the final enriched bead count going into the sequencing reaction.

If your read length distribution is strange,...
Forum: 454 Pyrosequencing 06-12-2009, 07:23 AM
Replies: 14
Views: 4,120
Posted By boss_hoss
hlu, what do you mean by primer dimers in...

hlu,

what do you mean by primer dimers in the sequencing reaction? wouldnt these be removed by SPRI during LC?
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