We have recently upgraded to the FLX+. We have had some successful quads with amplicons in the 600 to 800 base range. However, one quad with 16 samples shows nearly 100% key pass failure! It had a 20% enrichment similar to the quad beside it that was OK. The same barcode adaptors were used. The amplicons were amplified several cycles longer. By eye it is possible it might have more beads loaded than the other quad. Does anyone know what factors can cause such a large number of wells to have key pass failures?The control beads are proportionally low as well.
Seqanswers Leaderboard Ad
Collapse
Announcement
Collapse
No announcement yet.
X
-
Can you look through the PTP images for each of the early cycles by eye using gsRunBrowser to see if the key bases are there? Most sample keys are not compatible -- so if the library somehow got a little of a different sample key mixed in, it might cause issues.
--
Phillip
Comment
-
We had a breakthrough on this Friday afternoon. Although we thought it was a problem with our samples in that Quad at first, after support looked at it, they were certain it was not analyzed properly. I had mentioned some error messages I had had when discussing this problem earlier but they didnt think it was the problem. NOW it probably is! I am thinking that this FLX+ data could be overwhelming the processing of the data. I guess the biggest hint was poor control beads as well.
Comment
Latest Articles
Collapse
-
by seqadmin
Metagenomics has improved the way researchers study microorganisms across diverse environments. Historically, studying microorganisms relied on culturing them in the lab, a method that limits the investigation of many species since most are unculturable1. Metagenomics overcomes these issues by allowing the study of microorganisms regardless of their ability to be cultured or the environments they inhabit. Over time, the field has evolved, especially with the advent...-
Channel: Articles
09-23-2024, 06:35 AM -
-
by seqadmin
During the COVID-19 pandemic, scientists observed that while some individuals experienced severe illness when infected with SARS-CoV-2, others were barely affected. These disparities left researchers and clinicians wondering what causes the wide variations in response to viral infections and what role genetics plays.
Jean-Laurent Casanova, M.D., Ph.D., Professor at Rockefeller University, is a leading expert in this crossover between genetics and infectious...-
Channel: Articles
09-09-2024, 10:59 AM -
ad_right_rmr
Collapse
News
Collapse
Topics | Statistics | Last Post | ||
---|---|---|---|---|
Started by seqadmin, 10-02-2024, 04:51 AM
|
0 responses
13 views
0 likes
|
Last Post
by seqadmin
10-02-2024, 04:51 AM
|
||
Started by seqadmin, 10-01-2024, 07:10 AM
|
0 responses
21 views
0 likes
|
Last Post
by seqadmin
10-01-2024, 07:10 AM
|
||
Started by seqadmin, 09-30-2024, 08:33 AM
|
0 responses
25 views
0 likes
|
Last Post
by seqadmin
09-30-2024, 08:33 AM
|
||
Started by seqadmin, 09-26-2024, 12:57 PM
|
0 responses
18 views
0 likes
|
Last Post
by seqadmin
09-26-2024, 12:57 PM
|
Comment