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

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
ampure beads sc10021 Sample Prep / Library Generation 12 07-03-2017 02:26 AM
Ampure XP beads Etherella Illumina/Solexa 4 10-16-2013 06:05 AM
Ampure XP beads cleanup and concentration seqgirl123 Sample Prep / Library Generation 2 04-24-2013 09:40 AM
AMPure, amplicons and GS Jr enrichment beads suefo 454 Pyrosequencing 17 02-06-2012 04:24 AM

Thread Tools
Old 05-08-2014, 11:34 PM   #1
Jafar Jabbari
Location: Melbourne

Join Date: Jan 2013
Posts: 1,238
Default Freshness of Ethanol used for washing AMPure beads

Most of Illumina library prep protocols which uses AMPure beads for clean up or size selection recommends using freshly prepared 70% or 80% Ethanol for wash step. Both works very well and I have not noticed any difference in performance of eluted libraries washed by either. I wonder how often or how fresh any one else prepares their wash Ethanol. Fresh prep in literature is justified by presumed or real changes in ethanol concentration by absorption of air moisture. Considering that two recommended wash solutions differs by 10% in Ethanol content does freshness really makes any difference.
nucacidhunter is offline   Reply With Quote
Old 05-09-2014, 01:57 AM   #2
Senior Member
Location: Sweden

Join Date: Mar 2008
Posts: 324

It does not need to be made fresh. Possibly altered EtOH concentration affects washing but you will not lose DNA unless EtOH is below 40 % or so (I have not tested this myself but it can easily be done with a simple titration using a ladder).
Chipper is offline   Reply With Quote
Old 05-19-2014, 01:53 PM   #3
Location: Missoula, MT

Join Date: Jan 2014
Posts: 19

I have recently spoken to a few technicians and post doc about this. Everyone I know makes fresh ethanol for AMPure bead clean ups or size selection - some make 73%, others 74%, 75%, etc. (exact number seems to just be personal preference).

Found this, supporting what Chipper said: "The original AMPure protocol recommended use of a 70% ethanol wash solution. This needed to be freshly made, as over time it becomes more dilute since it absorbs atmospheric water and ethanol evaporates. When you rinse with more dilute ethanol more DNA gets washed away. We have found that this can be avoided by using an 80% ethanol solution instead. This washes just as well but does not need to be freshly prepared each time."
kzarn 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 04:05 PM.

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