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

Similar Threads
Thread Thread Starter Forum Replies Last Post
AMPure XP vs SPRI beads Tali7 Sample Prep / Library Generation 19 07-12-2017 12:13 PM
Help with TruSeq gDNA library prep BTS Illumina/Solexa 45 02-22-2017 11:09 AM
TruSeq or Nextera kit for library prep? rooty Sample Prep / Library Generation 12 10-29-2013 07:02 AM
Automation for TruSeq library prep? jlove Sample Prep / Library Generation 10 01-10-2012 09:34 AM
TruSeq RNA initial concentration and Ampure beads OnUrMark Sample Prep / Library Generation 3 09-27-2011 09:37 AM

Thread Tools
Old 08-09-2012, 05:19 AM   #1
Junior Member
Location: Gaithersburg, Md

Join Date: Apr 2012
Posts: 1
Default TruSeq with AmPure Beads in during Library Prep

Hi Everyone:

We are having some issues during library prep recently and are asking for any opinions and some help out there from our fellow Illumina users. I have been doing TruSeq sample prep now for a year and have had no real issues until lately. We have started to process anywhere from 60 to 92 samples for full Exome capture. We sheer by covaris and have a starting gDNA yield of >1ug so we have enough starting material even after viewing the samples with a HS Caliper Chip. We have tried changing all of our reagents and solutions to make sure everything has been fresh and nothing expired and are still seeing a dramatic drop off after the first LM-PCR. We are using NimbleGen V3 for our capture and using NEB Phusion Master Mix for the LM-PCR. So my biggest question is that it looks like we are losing the product during the AmPure steps and was wondering if anyone leaves the beads in during the TruSeq prep and then using the Qiagen Qiaquick clean up method to get rid of the beads before capture.

Sorry about the long explanation but any help would be greatly appreciated.
Malasky53 is offline   Reply With Quote
Old 08-09-2012, 08:08 AM   #2
Location: cambridge

Join Date: Apr 2010
Posts: 68

I haven't been involved in next gen for almost a year but I do remember ampure beads can go off. Have you tried a new fresh aliquot of your beads? Also you could try and ph you beads and then contact your ampure rep to see if there is a problem there.
niceday is offline   Reply With Quote

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 07:48 PM.

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