Hi All,
I have an unusual problem with the consistency of emPCR kits that were received two months ago (emPCR oil and breaking kit (Lot: 93896820), emPCR Lib-L kit (lot: 93896420) and Bead recovery kit (lot: 93893620). Of the eight kits, three worked well but five emPCRs have failed.
We calculate the copy number using the library concentration from Qubit and average size from the bioanalyzer, similar to what Roche suggested for quantification of pair-end libraries. This method has worked well in past as it has worked for three emPCRs from this batch. We also run a normal PCR reaction to test the library amplificability which suggest that the libraries are fine.
I feel the problem is with the emPCR kits because:
1. We had a gloppy thing floating over the beads during the bead recovery three times it failed. It appears to be the problem with emulsion oil.
2. I repeated the emPCR for the library that has worked before (using same amount) but it failed the second time.
3. I tripled the amount of library in the emPCR (as suggested in the emPCR manual) but still not enough beads were recovered.
I was wondering if someone else experienced similar issue. Any suggestions will be highly appreciated.
Thanks and regards,
Seqanswers Leaderboard Ad
Collapse
Announcement
Collapse
No announcement yet.
X
Latest Articles
Collapse
-
by seqadmin
The field of epigenetics has traditionally concentrated more on DNA and how changes like methylation and phosphorylation of histones impact gene expression and regulation. However, our increased understanding of RNA modifications and their importance in cellular processes has led to a rise in epitranscriptomics research. “Epitranscriptomics brings together the concepts of epigenetics and gene expression,” explained Adrien Leger, PhD, Principal Research Scientist...-
Channel: Articles
04-22-2024, 07:01 AM -
-
by seqadmin
Proteins are often described as the workhorses of the cell, and identifying their sequences is key to understanding their role in biological processes and disease. Currently, the most common technique used to determine protein sequences is mass spectrometry. While still a valuable tool, mass spectrometry faces several limitations and requires a highly experienced scientist familiar with the equipment to operate it. Additionally, other proteomic methods, like affinity assays, are constrained...-
Channel: Articles
04-04-2024, 04:25 PM -
ad_right_rmr
Collapse
News
Collapse
Topics | Statistics | Last Post | ||
---|---|---|---|---|
Started by seqadmin, Yesterday, 11:49 AM
|
0 responses
15 views
0 likes
|
Last Post
by seqadmin
Yesterday, 11:49 AM
|
||
Started by seqadmin, 04-24-2024, 08:47 AM
|
0 responses
16 views
0 likes
|
Last Post
by seqadmin
04-24-2024, 08:47 AM
|
||
Started by seqadmin, 04-11-2024, 12:08 PM
|
0 responses
61 views
0 likes
|
Last Post
by seqadmin
04-11-2024, 12:08 PM
|
||
Started by seqadmin, 04-10-2024, 10:19 PM
|
0 responses
60 views
0 likes
|
Last Post
by seqadmin
04-10-2024, 10:19 PM
|