SEQanswers

Go Back   SEQanswers > Applications Forums > Sample Prep / Library Generation



Similar Threads
Thread Thread Starter Forum Replies Last Post
what specific sequence is used for annealing to flow cell? ein_io Illumina/Solexa 3 12-13-2011 07:56 AM
sequencing primer annealing Isabel_Bodmer 454 Pyrosequencing 2 11-18-2011 12:20 PM
Current read1, index, and read2 primers for HiSeq2000 SeqVicious Illumina/Solexa 1 09-26-2011 02:12 PM
Illumina Pair end primers and adaptors for multiplexing mimi_lupton Sample Prep / Library Generation 1 05-25-2011 07:57 AM
Annealing index and primers for multiplexing alisrpp Illumina/Solexa 0 02-22-2011 05:12 PM

Reply
 
Thread Tools
Old 02-23-2011, 08:05 AM   #1
alisrpp
Member
 
Location: New York

Join Date: Dec 2010
Posts: 40
Default Annealing index and primers for multiplexing??

Hi,
We are planning to do a multiplexed cDNA library to run on Illumina . We bought the indexes and primers from IDT (sequences from Illumina) but we have some questions. We were reading that Illumina changed the protocol and now it uses the index+PCRprimer (all together) instead of using the index as a "third" primer.
We were thinking about annealing the index+primer and use it as a single one but we are not sure if it is gonna work. Does anybody have tried that?
If so, which temperature?
Otherwise we will use as separated things. Has anybody got good results with this?

Thanks
alisrpp is offline   Reply With Quote
Old 03-09-2011, 07:51 PM   #2
RNAseqer
Member
 
Location: London

Join Date: Sep 2010
Posts: 22
Default

ahhh...we used to do this but not anymore. Just couldn't get good annealing, we'd see multiple peaks. I recommend using the TruSeq adapters from Illumina or the DNA Adapters from Bioo which are sold separately from their library prep kit.
RNAseqer is offline   Reply With Quote
Reply

Tags
annealing, illumina, index, multiplexing, primer

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 02:55 PM.


Powered by vBulletin® Version 3.8.9
Copyright ©2000 - 2019, vBulletin Solutions, Inc.
Single Sign On provided by vBSSO