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Forum: 454 Pyrosequencing 07-11-2013, 08:05 AM
Replies: 2
Views: 2,284
Posted By MEJ
Amplicon sequencing - fusion primers ordering

Hi,

The phosphorothioate bases are there to inhibit any exonuclease activity, so you can order them without. It did work in our hands but if you get any contamination you'll have a problem!

The...
Forum: 454 Pyrosequencing 07-11-2013, 07:53 AM
Replies: 1
Views: 1,500
Posted By MEJ
A coverage depth of 15-20x in our hands on a FLX+...

A coverage depth of 15-20x in our hands on a FLX+ is more then enough for a 2Mb bacterial genome. More coverage didn't lead to less contigs.
Forum: 454 Pyrosequencing 11-22-2010, 03:47 AM
Replies: 11
Views: 3,929
Posted By MEJ
Thanks you all for replying! Our emPCR problems...

Thanks you all for replying!
Our emPCR problems started with the new oil. I can't really tell what goes wrong since we sometimes get good results and somtimes it's crap even with the same lot...
Forum: 454 Pyrosequencing 11-08-2010, 06:40 AM
Replies: 11
Views: 3,929
Posted By MEJ
emPCR

In August we encountered a problem with the delivery of emPCR kits, the oil was kept in back order and could only be delivered after a month. In the kit part with the oil a yellow note is included:...
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