SEQanswers

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



Similar Threads
Thread Thread Starter Forum Replies Last Post
How to mix single index and dual index TruSeq libraries pmiguel Illumina/Solexa 31 05-24-2017 03:10 AM
Pooling TruSeq (single index) and Nextera XT (dual index) in one MiSeq run? Carosmile Sample Prep / Library Generation 1 05-16-2017 01:13 PM
MiSeq Run, Problem with Index reads spf2 Illumina/Solexa 6 08-24-2015 06:59 AM
Dual-index vs single index Ingeneious Illumina/Solexa 4 01-19-2015 11:22 AM
Mixing dual and single index TruSeq samples in a single MiSeq run pmiguel Illumina/Solexa 1 12-21-2012 05:21 AM

Reply
 
Thread Tools
Old 12-20-2017, 08:33 AM   #1
Min Jung Lee
Member
 
Location: Seoul Korea

Join Date: Jan 2014
Posts: 11
Default Miseq dual index Read1 (i7 index) problem?

During miseq run, good quaility of read1 and read3 (i5), read4.
But read2 i7 index is low quality.
I already found many posting about read3 (i5 index) problem.
but I can’t find about i7 index failure.
I use Truseq HT dual index primer for 5 sample.
Help me please.

Last edited by Min Jung Lee; 12-20-2017 at 08:36 AM.
Min Jung Lee is offline   Reply With Quote
Old 12-20-2017, 05:20 PM   #2
nucacidhunter
Jafar Jabbari
 
Location: Melbourne

Join Date: Jan 2013
Posts: 1,179
Default

It could be that i7 indexes are not diverse enough. You can check Illumina pooling guidelines for duel indexed adapters (look at page 21).

http://sapac.support.illumina.com/co...5042173-01.pdf
nucacidhunter is offline   Reply With Quote
Old 12-20-2017, 05:24 PM   #3
Min Jung Lee
Member
 
Location: Seoul Korea

Join Date: Jan 2014
Posts: 11
Default

Thank you for your reply.
but, my library is enough diversity.
Min Jung Lee is offline   Reply With Quote
Old 12-20-2017, 07:28 PM   #4
nucacidhunter
Jafar Jabbari
 
Location: Melbourne

Join Date: Jan 2013
Posts: 1,179
Default

I am pointing to index diversity not the library. I wonder which wells from adapter plate you have used.
nucacidhunter is offline   Reply With Quote
Old 01-08-2018, 11:22 AM   #5
pmiguel
Senior Member
 
Location: Purdue University, West Lafayette, Indiana

Join Date: Aug 2008
Posts: 2,291
Default

What sort of adapters are these that you are using?
Illumina instruments that prime i5 using the flow cell oligos (including the MiSeq) will always produce i5 sequence from a library molecule. But to produce an i7 read you need one of the i7 indexing primers to anneal in your amplicon. MiSeq's seem to have a fairly high annealing temp for their sequencing primers...

--
Phillip
pmiguel 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 08:27 PM.


Powered by vBulletin® Version 3.8.9
Copyright ©2000 - 2018, vBulletin Solutions, Inc.
Single Sign On provided by vBSSO