SEQanswers

Go Back   SEQanswers > Applications Forums > RNA Sequencing



Similar Threads
Thread Thread Starter Forum Replies Last Post
miRNA Extraction from Plasma DrDTonge Sample Prep / Library Generation 8 08-11-2014 06:27 AM
RNA Extraction from Sludge megalodon Sample Prep / Library Generation 4 04-01-2011 05:54 PM
Gene extraction software andreitudor Bioinformatics 1 03-16-2011 01:21 PM
reg fastq extraction reena_narayanan General 3 03-13-2011 11:09 PM
templiphi dna extraction seqgirl123 General 0 10-26-2008 07:50 PM

Reply
 
Thread Tools
Old 05-05-2011, 02:59 PM   #1
Dan_cham
Member
 
Location: Israel

Join Date: Feb 2011
Posts: 13
Default RNA extraction from lizard

Hello all

I want to sequence the transcriptome of a new organism (species of lizard)

i order to do that, i need first to extract rna from fresh tissue. i follow protocol but I got an rna result that suggesting that my sample was degredated. but because I don't know many people who try to extract rna from reptiles i dont know if maybe the sample suppose to be different from standard extraction from mammals.

attached please find a pdf version of bio analyzer and gel electrophoresis of my samples (samples 11 and 12)

will be happy to hear ideas

Dan
Attached Files
File Type: pdf Next-generation DNA sequencing of paired-end tags.pdf (1.39 MB, 31 views)
Dan_cham is offline   Reply With Quote
Old 05-05-2011, 03:39 PM   #2
Wallysb01
Senior Member
 
Location: San Francisco, CA

Join Date: Feb 2011
Posts: 286
Default

We've used the mirVana kit from Ambion on lizard tissues and had no real problems and don't see any differences in the prep with mammals. Usually RNA degradation can be avoid by just processing the samples quickly. Dissecting and getting into cold extraction/lysis buffer quickly is key. Then flash freeze for storage or proceed quickly through the rest of the protocol.
Wallysb01 is offline   Reply With Quote
Old 05-05-2011, 06:02 PM   #3
ECO
--Site Admin--
 
Location: SF Bay Area, CA, USA

Join Date: Oct 2007
Posts: 1,358
Default

Uh....you attached a PDF of a Ruan RNA-seq paper...just FYI.

Flash freeze, grind, into Tri reagent of choice.
ECO is offline   Reply With Quote
Reply

Tags
rna extraction

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 07:47 PM.


Powered by vBulletin® Version 3.8.9
Copyright ©2000 - 2019, vBulletin Solutions, Inc.
Single Sign On provided by vBSSO