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Forum: RNA Sequencing 09-27-2009, 06:02 PM
Replies: 36
Views: 22,309
Posted By wiggy
My sequences were on ncbi so I just used it to...

My sequences were on ncbi so I just used it to design primers.

Or you could find highly conserved regions, create primers, sequence, and repeat until you get the whole sequence.

I believe this...
Forum: 454 Pyrosequencing 08-07-2009, 07:23 AM
Replies: 6
Views: 3,507
Posted By wiggy
We are using random ninemers after poly A...

We are using random ninemers after poly A purification. We are seeing large 2-3 KB transcripts, some transcribed at low levels, making it into the first strand using this method. Our qRTPCR primers...
Forum: RNA Sequencing 08-01-2009, 02:16 PM
Replies: 36
Views: 22,309
Posted By wiggy
We subjected RNA to 2 rounds of PolyA...

We subjected RNA to 2 rounds of PolyA purification from Promega and also to 1 round of Ribominus.

After analyzing via qRTPCR for both 18S and 28S rRNA we found the PolyA kit almost completely...
Forum: 454 Pyrosequencing 06-18-2009, 11:11 AM
Replies: 0
Views: 2,674
Posted By wiggy
Size Selection Tricks??

Hello everyone this is my first post.

We are going to be running about 4 different EST libraries through the |Roche 454, and are trying to save some money and time by size selecting the fragments...
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