Oligo dT VN is used for anchored cDNA synthesis. But what does "VN" in the end do? Please help. Thanks.
Seqanswers Leaderboard Ad
Collapse
Announcement
Collapse
No announcement yet.
X
-
Oligo dT length is normally 30nt but transcript poly A tail could be longer. Including VN at 3’ end of RT primer enriches for non-poly A region of transcripts and without it RT can be primed in poly A region so cDNA would have A/T homopolymer which in most sequencing applications is undesirable. A side effect of VN inclusion is possible bias in some transcripts conversion to cDNA.
-
why VN, why not just V for oligo dT anchor?
Why does almost everyone use 3'-VN as the anchor for oligo dT primers rather than using 3'-V?
The former only has a 1/12 chance of matching a given mRNA, vs. 1/3 for the latter. Plus 1/4 of the former will have T as the N, meaning there's a greater potential for annealing and extension within the polyA tail with VN vs V.
Only reason I can think of is legacy.
Thanks!
Mike
Comment
Latest Articles
Collapse
-
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 -
-
by seqadmin
The first FDA-approved CRISPR-based therapy marked the transition of therapeutic gene editing from a dream to reality1. CRISPR technologies have streamlined gene editing, and CRISPR screens have become an important approach for identifying genes involved in disease processes2. This technique introduces targeted mutations across numerous genes, enabling large-scale identification of gene functions, interactions, and pathways3. Identifying the full range...-
Channel: Articles
08-27-2024, 04:44 AM -
ad_right_rmr
Collapse
News
Collapse
Topics | Statistics | Last Post | ||
---|---|---|---|---|
Started by seqadmin, Today, 06:25 AM
|
0 responses
13 views
0 likes
|
Last Post
by seqadmin
Today, 06:25 AM
|
||
Started by seqadmin, Yesterday, 01:02 PM
|
0 responses
12 views
0 likes
|
Last Post
by seqadmin
Yesterday, 01:02 PM
|
||
Started by seqadmin, 09-18-2024, 06:39 AM
|
0 responses
14 views
0 likes
|
Last Post
by seqadmin
09-18-2024, 06:39 AM
|
||
Started by seqadmin, 09-11-2024, 02:44 PM
|
0 responses
14 views
0 likes
|
Last Post
by seqadmin
09-11-2024, 02:44 PM
|
Comment