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

Similar Threads
Thread Thread Starter Forum Replies Last Post
Single Cell RNA-seq Troubleshooting tmm343 Sample Prep / Library Generation 6 10-17-2016 01:34 PM
Neoprep RNA-Seq troubleshooting brcseq Sample Prep / Library Generation 2 09-29-2016 04:04 PM
KAPA qPCR troubleshooting lorendarith Illumina/Solexa 2 02-12-2014 04:10 AM
Me-DIP-seq troubleshooting, several questions NearyJL78 Illumina/Solexa 7 09-27-2011 09:24 AM
troubleshooting illumina seq biochembug Illumina/Solexa 0 03-04-2011 09:06 AM

Thread Tools
Old 09-22-2017, 07:04 AM   #1
Junior Member
Location: Manchester, UK

Join Date: Sep 2017
Posts: 1
Default ATAQ-Seq qPCR troubleshooting


I am attempting to perform ATAQ-seq on human macrophages. I have performed the protocol, though have encountered an issue at the qPCR step (after the initial 5 cycle amplification step). I find i have no amplification at all.

I was worried it was because i lost my cell pellet during the initial wash stages so i have nanodropped my samples at various steps:

1. After the initial purification following the transposition reaction
2. After the initial 5 cycle PCR amplification step
3. After final purification step

However, i have DNA at all of these steps, indicating i haven't lost my pellet and the DNA is being amplified.

For this protocol i am using the Illumina Nextera Index Kit (FC-121-1012) as the primers.
Regarding this i have two questions:
1. Are these indices compatible with qPCR?
2. For each sample should use the same index pair used in the amplification PCR in the qPCR?

Any other suggestions would be greatly appreciated!
d123morg is offline   Reply With Quote

atac-seq, atacseq, illumina, nextera indices, qpcr

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 02:02 PM.

Powered by vBulletin® Version 3.8.9
Copyright ©2000 - 2020, vBulletin Solutions, Inc.
Single Sign On provided by vBSSO