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

Similar Threads
Thread Thread Starter Forum Replies Last Post
PippinPrep for TruSeq small RNA library selection visserm Sample Prep / Library Generation 3 09-26-2013 09:36 AM
Small RNA library preparation nirwalniraj Sample Prep / Library Generation 5 02-01-2012 09:42 AM
small RNA Truseq library sequencing primers deepakpatilp Illumina/Solexa 0 08-26-2011 02:22 AM
Problem with small RNA library preparation bpa400 Illumina/Solexa 3 11-01-2010 02:56 PM

Thread Tools
Old 02-15-2013, 02:32 AM   #1
Ilena Li Mura
Junior Member
Location: Padova

Join Date: Feb 2013
Posts: 2
Default Illumina TruSeq Small RNA library preparation

Hi, I'm some problems with micro RNA library preparation using Illumina TruSeq Small RNA protocol.
I attach the images of bioanalyzer before gel purification of Ambion Human Brain Total RNA that I use as positive control.
One images is about the control after 11 cycles of PCR Amplification (that is the cycles number that I usually use), but as you can see there is an evident decrease of material greater than approximately 100bp. We observe this kind of profile in all our samples and, in our experience, these are not normal patterns.
The other image is the same control with 15 cycles of PCR amplification and is quite different.

Are these patterns normal or there is any trouble?

Have you any suggestion to solve the problems?

Thanks in advance
Attached Images
File Type: png 11cycles.png (124.7 KB, 74 views)
File Type: png 15cycles.png (124.7 KB, 63 views)
Ilena Li Mura is offline   Reply With Quote
Old 06-09-2018, 01:42 AM   #2
Junior Member
Location: India

Join Date: Jun 2015
Posts: 1

At 15 cycles, 147bp and 198bp bands reduced and 137bp and 184bp bands emerged. How did the gel purification go? On the gel also did you see similar pattern? Were you able to cutout the band excluding adapter dimer. However, i dont see adpter dimer band in this case. Thanks.
psreddy 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 04:45 PM.

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