Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > General Science Forums > Biochemistry Forum > RNA Forum
Register Search Today's Posts Mark Forums Read

RNA Forum RNA Forum. Discuss RNA including mRNA structure and function, rRNA, tRNAs, and the biochemistry of other RNA molecules.


RNA purification from reticulocytes

RNA purification from reticulocytes - RNA Forum

RNA purification from reticulocytes - RNA Forum. Discuss RNA including mRNA structure and function, rRNA, tRNAs, and the biochemistry of other RNA molecules.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 08-01-2011, 02:04 PM
Pipette Filler
Points: 1,355, Level: 21 Points: 1,355, Level: 21 Points: 1,355, Level: 21
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jul 2011
Location: Argentine
Posts: 18
Thanks: 1
Thanked 0 Times in 0 Posts
Default RNA purification from reticulocytes



Hi, does anyone have the protocol to extract RNA from reticulocytes?thanks!!!
Reply With Quote
  #2  
Old 02-12-2013, 03:56 PM
Summer Student
Points: 240, Level: 4 Points: 240, Level: 4 Points: 240, Level: 4
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Feb 2013
Posts: 56
Thanks: 0
Thanked 0 Times in 0 Posts
Default エアジョーダン 通販

[

[

[
Reply With Quote
  #3  
Old 03-17-2014, 04:04 PM
Zagami's Avatar
Ph.D Doctorate
Points: 4,035, Level: 42 Points: 4,035, Level: 42 Points: 4,035, Level: 42
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Sep 2009
Location: Tabaka Mission Hospital - Kiisi -Kenya & Italy Sicily
Posts: 159
Thanks: 2
Thanked 28 Times in 24 Posts
Default Re: RNA purification from reticulocytes

Adult human reticulocytes are isolated from 20 ml of peripheral blood in heparin following a process of leukodepletion. Leukocytes are concentrated by centrifugation at 3500 rpm for 30 min at 4 C, forming a disc (buffy coat) at the plasma-red cell interface. Both the plasma and buffy coat are removed, and the cells are washed three times with 10 ml of ice-cold PBS. At the third wash, reticulocytes are concentrated by centrifugation at 3500 rpm for 30 min at 4 C. The supernatant is removed, and the top 700 μl reticulocyte-rich fraction is added to 4 ml of ice-cold PBS. This red cell suspension is layered on top of a column consisting of about 4.5 ml of 2 parts cellulose (Sigma cod. C8002) and 1 part sigmacell type 50 microcrystallin cellulose (Sigma cod. S5504) in normal saline. The red cells are eluted by gently centrifuging the column at 1000 rpm at 4 C for 1 min. This leukodepleted eluate (∼2.5 ml) is washed three times in 10 ml of ice-cold PBS. Finally, the reticulocyte-enriched eluted cells are recovered by centrifugation and resuspended in 2 ml of ice-cold PBS. RNA is extracted with 5 ml TRI reagent (Sigma cod. T9424).
Reply With Quote
Reply

Tags
purification , reticulocytes , rna


« ESEfinder | - »
Thread Tools
Display Modes

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
Trackbacks are On
Pingbacks are On
Refbacks are On

Forum Jump

Similar Threads
Thread Thread Starter Forum Replies Last Post
site-directed mutagenesis - primer purification majihec Molecular Biology Techniques 0 05-02-2011 02:17 PM
His-Tag purification - contaminated protein Elayoe Recombinant Protein Forum 3 09-29-2009 09:10 PM
His-tag protein purification under denaturing conditions Arabidop Molecular Biology Articles and Protocols 5 06-28-2009 06:05 AM
IgM purification Anteros Antibody Forum 2 07-13-2008 09:46 AM
inactive enzyme after purification with Ni-TED piefke Recombinant Protein Forum 0 05-02-2008 08:14 AM


All times are GMT. The time now is 11:35 PM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.12261 seconds with 16 queries