SEQanswers

Go Back   SEQanswers > Sequencing Technologies/Companies > Illumina/Solexa



Similar Threads
Thread Thread Starter Forum Replies Last Post
polyA containing reads in RNA-seq wenhuang Bioinformatics 6 02-19-2015 04:16 PM
polyA selection kits chaos81 Sample Prep / Library Generation 8 04-11-2012 09:55 AM
DSN vs polyA protocol publications? Jeltje RNA Sequencing 6 08-14-2011 12:48 AM
PolyA reads are disappeared? SOLiDrobino SOLiD 5 08-09-2011 06:24 AM
polyA+ yields from total RNA? pmiguel RNA Sequencing 10 04-28-2010 05:47 AM

Reply
 
Thread Tools
Old 09-22-2011, 05:56 AM   #1
biochembug
Member
 
Location: New York

Join Date: Mar 2011
Posts: 26
Default PolyA on flowcell

Hi Friends

If I plan to clone RNA near its 3' end and somehow select just the 3' ends. What do you think about clusters when sequencing the A tail. I am planning to sequence from the back-end of the RNA. Will the clusters be unique or because every spot will glow for A (or T) it might confuse with the clusters?

Does anyone have idea whether we can place a barcode before the insert and read it as a part of insert? Any alternatives?

Please share your experiences and suggest some other ideas?

Biochembug
biochembug is offline   Reply With Quote
Old 09-22-2011, 09:05 AM   #2
HESmith
Senior Member
 
Location: Bethesda MD

Join Date: Oct 2009
Posts: 509
Default

Accurate cluster calling requires sequence diversity during the first 4-5 cycles. You can design your primers to sequence from the non-A end, or engineer 5mer barcodes with equal representation before the polyA.
HESmith is offline   Reply With Quote
Reply

Thread Tools

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off




All times are GMT -8. The time now is 05:32 AM.


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