SEQanswers

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



Similar Threads
Thread Thread Starter Forum Replies Last Post
Custom primer failed kal33na Illumina/Solexa 1 04-22-2013 02:39 PM
50+% of my HiSEQ reads are 3' primer (custom primer used) ZAAB Sample Prep / Library Generation 3 02-06-2012 12:00 PM
Can smRNA sequencing primer and genomic DNA sequencing primer mixed in one lane? whimsy Illumina/Solexa 3 04-06-2011 10:30 AM
custom primer menglv Illumina/Solexa 3 01-18-2011 02:01 PM
PubMed: Systematic cross-validation of 454 sequencing and pyrosequencing for the exac Newsbot! Literature Watch 0 01-18-2011 03:00 AM

Reply
 
Thread Tools
Old 07-20-2012, 12:28 AM   #1
Akira
Member
 
Location: Malaysia

Join Date: Sep 2010
Posts: 16
Default Validation of Custom Sequencing Primer

Hi.

I would like to use my own sequencing primer. I design it with properties (Tm
and GC content and length etc) close to standard Illumina Sequencing Primer.

To check its performance I set up a PCR reaction with the primer as forward and P7 primer as reverse. But I don't know what is the annealing temperature I should use.

Anyone know what is the temperature for Illumina Sequencing Primer to anneal during cBOT last step (not sure if it's correct)? So that I can follow the temperature in my PCR step to see if my primer can anneal.

Please help. Thanks

- NGS newbie.
Akira is offline   Reply With Quote
Old 11-05-2012, 10:22 AM   #2
giampe
Member
 
Location: Bari, Italy

Join Date: Aug 2009
Posts: 21
Default

Hi,
maybe if you take a look into http://molbiol.ru/wiki/Cluster_generation_on_cBot
probably it will help to answer your question.
giampe is offline   Reply With Quote
Reply

Tags
annealing temperature, cbot, custom, illumina, sequencing 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 02:22 AM.


Powered by vBulletin® Version 3.8.9
Copyright ©2000 - 2017, vBulletin Solutions, Inc.
Single Sign On provided by vBSSO