SEQanswers

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



Similar Threads
Thread Thread Starter Forum Replies Last Post
New Ribo-Zero Gold Kit (Human/Mouse/Rat) epibio Vendor Forum 7 08-01-2014 01:35 PM
Magnetic bead washers Benny Chain Illumina/Solexa 5 10-09-2013 02:29 PM
New Ribo-Zero™ Kit (Epidemiology):rRNA removal from microbial-infected mammalian RNA epibio Vendor Forum 0 12-12-2012 02:24 PM
PubMed: Magnetic sequencing. Newsbot! Literature Watch 2 09-26-2012 01:05 AM
Ribo-Zero (Human/Mouse/Rat) kit now in magnetic format epibio Vendor Forum 1 02-06-2012 07:48 AM

Reply
 
Thread Tools
Old 05-14-2014, 06:47 AM   #1
marRoma
Junior Member
 
Location: roma

Join Date: Jan 2014
Posts: 6
Default ribo-zero magnetic kit

hi!

I have a question about the way to purify the rRna-depleted RNA , after the ribo-zero magnetic kit. the ribozero kit suggests either the ethanol precipitation or the rneasy cleanup kit or agenancourt RNAclean but i have the rna clean and concentrator kit by zymo research; can i use it istead of that one suggested by epicentre? does anyone have experience with this kit? because i'd like to avoid the ethanol precipitation, i've read that the concentration of RNA decreases a lot.

thank you

Margherita
marRoma is offline   Reply With Quote
Old 05-15-2014, 04:42 AM   #2
nucacidhunter
Jafar Jabbari
 
Location: Melbourne

Join Date: Jan 2013
Posts: 1,238
Default

We use RNAClean knowing that it is poor in recovering small RNA fraction which we are not interested. Methods recommended by Epicentre have been tested for various properties. One can use similar product, but it is unknown if they introduce any bias. Column based cleaning products normally have different size bias (some are poor in recovering large fragments) which may or may not be crucial for downstream applications. So, it is best to use recommended products to be safe.
nucacidhunter is offline   Reply With Quote
Old 05-15-2014, 05:22 PM   #3
kerplunk412
Senior Member
 
Location: Bioo Scientific, Austin, TX, USA

Join Date: Jun 2012
Posts: 119
Default

I actually tested RNA cleanup methods about a year and a half ago. Ethanol precipitation was the highest yielding method, but it does take a long time and there is a lot of chance for loss when dealing with tiny pellets. The Zymo C&C worked well in my hands, and I even changed the lab RNA-Seq protocol to use it instead of RNEasy Minelute. If you use the RNEasy kit it is very important that you use the Ribozero protocol, or more specifically the amount of ethanol indicated in the Ribozero protocol. In my experience the yield from the Minelute kit is not very good unless you use much more ethanol than indicated in the manual. The Zymo C&C kit worked well with the volume of ethanol indicated in its manual, but I do recommend re-applying the eluate to the column at least once for increased yield, especially if eluting in a small volume.
kerplunk412 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 01:20 PM.


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