SEQanswers

Go Back   SEQanswers > Search Forums


Showing results 1 to 9 of 9
Search took 0.00 seconds.
Search: Posts Made By: eladSeq
Forum: 454 Pyrosequencing 11-28-2012, 02:44 PM
Replies: 4
Views: 3,197
Posted By eladSeq
AJ, thanks for the feedback. Our provider did...

AJ, thanks for the feedback. Our provider did also run the shotgun pipeline and that improved things a little bit to 10% pass filter and length actually increased from 355 to 400 bp. They also tried...
Forum: 454 Pyrosequencing 11-28-2012, 12:14 PM
Replies: 7
Views: 4,673
Posted By eladSeq
Hi, We are having trouble sequencing our...

Hi,
We are having trouble sequencing our amplicon pool due to short read failure causing very low pass filter %. For this library, I combined amplicons and then did an agarose gel extraction using a...
Forum: 454 Pyrosequencing 11-28-2012, 08:24 AM
Replies: 4
Views: 3,197
Posted By eladSeq
Low pass filter % due to short reads

Hi,
We previously performed a 454 amplicon sequencing run on our gene library with great results back in March '12. We prepared a new amplicon library of the same gene essentially the same way as...
Forum: 454 Pyrosequencing 03-30-2012, 05:19 PM
Replies: 2
Views: 4,271
Posted By eladSeq
PCR Jumping (Template Switching) in Amplicon sequencing

Hi,

We performed a full plate amplicon run on 454 FLX, on a gene library in which a single mutation was introduced to each molecule at a different position. We are observing a ~5X higher mutation...
Forum: 454 Pyrosequencing 03-03-2012, 10:49 AM
Replies: 4
Views: 9,908
Posted By eladSeq
Using the Lib-A emPCR kit, we are having an issue...

Using the Lib-A emPCR kit, we are having an issue where we observe good enrichment with the "B" beads, but no enrichment using the "A" beads. Since the amplicons each have an A and B adapter sequence...
Forum: 454 Pyrosequencing 01-16-2012, 02:58 PM
Replies: 19
Views: 6,613
Posted By eladSeq
So I have designed sets of A and B fusion primers...

So I have designed sets of A and B fusion primers with varying barcodes to get the desired offsetting of the flow reads. This will require 42 primers to make and pool 21 amplicons. Because of the...
Forum: Illumina/Solexa 12-23-2011, 08:30 AM
Replies: 3
Views: 4,551
Posted By eladSeq
Any determination on whether the fusion primers...

Any determination on whether the fusion primers need to be HPLC purified or just desalted?
Forum: 454 Pyrosequencing 12-23-2011, 08:00 AM
Replies: 19
Views: 6,613
Posted By eladSeq
Amplicon sequencing not supported on FLX+

Thank you everybody for your input.

Indeed, FLX+ is not currently supported for amplicons. From my understanding, there would be too much "flashing" due to the homogeneity of the sequences, such...
Forum: 454 Pyrosequencing 12-06-2011, 11:50 AM
Replies: 19
Views: 6,613
Posted By eladSeq
454 GS FLX+ long reads question

Hi,

I am a newcomer to deep sequencing and wanted to get some feedback on my project. My library consists of ~18,000 variants of an 860 bp gene on a plasmid. Because of its length I'm planning to...
Showing results 1 to 9 of 9

 


All times are GMT -8. The time now is 06:31 AM.


Powered by vBulletin® Version 3.8.9
Copyright ©2000 - 2021, vBulletin Solutions, Inc.
Single Sign On provided by vBSSO