Using the illumina mRNA Seq kits I created 8 libraries last week; they are completed through pcr enrichment and ready to run on the GAs. For the most part the traces looked good however on all 8 samples there is an odd peak at around 75 bps. When I performed a gel cut out at ~300 bps I don't believe I introduced any other material so I am not sure where the extra peak is coming from. It seems to me that it could be a primer dimer from pcr enrichment or some sort of contamination in the gel of my bioanalyzer kit, either way I don't think those would affect the reads on a GA but I'd like to hear some input from you guys. The scans are attached bellow
Seqanswers Leaderboard Ad
Collapse
Announcement
Collapse
No announcement yet.
X
-
-
If you used a TruSeq kit to construct these, then my guess is that the ~75 "bps" peak is actually the PCR primers used for enrichment PCR. These primers, from the "PPC" (PCR primer cocktail) tube give a peak in that region of the electropherogram of both DNA high sensitivity chips and RNA (pico) chips. See:
Any non-primary sequence heritable modification of genetic material. ChIP-SEQ, DNA methylation (Bisulfite-SEQ), chromatin modifications (methylation, acetylation, etc), non coding RNA.
We don't run the DNA1000 chips in our lab, so have never looked at it on that type of chip.
--
Phillip
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, 08:47 AM
|
0 responses
16 views
0 likes
|
Last Post
by seqadmin
Yesterday, 08:47 AM
|
||
Started by seqadmin, 04-11-2024, 12:08 PM
|
0 responses
60 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
|
||
Started by seqadmin, 04-10-2024, 09:21 AM
|
0 responses
54 views
0 likes
|
Last Post
by seqadmin
04-10-2024, 09:21 AM
|
Comment