Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > DNA Techniques > DNA Extraction Forum
Register Search Today's Posts Mark Forums Read

DNA Extraction Forum DNA Isolation Forum. A forum specifically about questions on DNA extraction and purification.


DNA extraction from buffy coat

DNA extraction from buffy coat - DNA Extraction Forum

DNA extraction from buffy coat - DNA Isolation Forum. A forum specifically about questions on DNA extraction and purification.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 08-17-2011, 02:59 AM
Pipette Filler
Points: 295, Level: 5 Points: 295, Level: 5 Points: 295, Level: 5
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jul 2011
Posts: 19
Thanks: 0
Thanked 2 Times in 2 Posts
Default DNA extraction from buffy coat



I'm looking to isolate DNA and eventually RNA from buffy coat. I receive whole blood in purple top vacutainers treated with EDTA. These tubes are centrifuged at 3000 rpm for 10 minutes to separate the whole blood into plasma buffy coat and rbc's. The plasma is drawn off with a transfer pipette to 2.0 ml microtubes and spun at 3000 rpm for 10 minutes to remove particulates, then transfered to cryovials and stored at -80.

I draw off the buffy coat and place it into a 15 ml conical tube filled with cold 10 mM tris 1mM EDTA. I let the tubes sit on ice for about 10 minutes then spin them at 3000 rpm for 10 minutes, pour off the supernatant and repeat with a few more 10 mL washes. A few rbc's persist but the majority are removed. The buffy coat is placed in a cryovial and stored at -80.

I'm using the fujifilm quickgene with various kits to isolate RNA and DNA from the frozen buffy coat.

I'm looking for suggestions on a more efficient method of obtaining buffy coat as well as extracting DNA.
Reply With Quote
Reply

Tags
buffy , buffy coat , centrifuge , coat , dna , extraction , extraction or isolation


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
buffy coat count moleculargeek21 DNA Forum 6 08-17-2011 04:01 PM
DNA from Buffy coat moleculargeek21 DNA Extraction Forum 0 08-17-2011 02:42 PM
Buffy coat moleculargeek21 DNA Forum 0 07-19-2011 04:26 PM
Extraction of whole blood DNA and RNA bioteke DNA Extraction Forum 0 07-07-2010 03:33 AM
RNA isolation from frozen buffy coat Anita Protocols and Methods Forum 0 07-03-2003 01:01 PM


All times are GMT. The time now is 09:52 AM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.11276 seconds with 16 queries