Go Back   Molecular Biology Forum > Molecular Research Topics Forum > Molecular Biology Techniques > Epigenetics Forum: DNA Methylation, Histone and Chromatin Study
Register Blogs FAQ Members List Calendar Science Groups New! Arcade Search Today's Posts Mark Forums Read

Epigenetics Forum: DNA Methylation, Histone and Chromatin Study An Epigenetics Forum Dealing with DNA Methylation, the study of histones and chromatin


Problem with extraction of DNA from Trizol

Epigenetics Forum: DNA Methylation, Histone and Chromatin Study

An Epigenetics Forum Dealing with DNA Methylation, the study of histones and chromatin



Register Molecular Biology Forums
Reply
 
LinkBack Thread Tools Display Modes
  #1 (permalink)  
Old 01-10-2008, 07:24 PM
Pipette Filler
Points: 201, Level: 4Points: 201, Level: 4Points: 201, Level: 4
Activity: 0%Activity: 0%Activity: 0%
 

Join Date: Jan 2008
Posts: 4
max_rempel RSS Feed
Default Problem with extraction of DNA from Trizol

I have generated a number of interesting mouse epidermis samples in Trizol. The treatments include the ones which may affect chronic state of the epidermis and therefore are interesting for methylation analysis. I was able to extract and use RNA from these samples. Now I am trying to extract DNA from the rest of each trizol sample.
The first try gave me very poor quality and low yields of DNA.

My QUESTION: has anyone extracted DNA from Trizol samples and what was the protocol?
Digg this Post!Add Post to del.icio.usBookmark Post in TechnoratiFurl this Post!Spurl this Post!Reddit!
Reply With Quote
Alt Today
Advertising
Google Adsense
 
This advertising will not be shown
in this way to registered members.
Register your free account today
and become a member on
Molecular Biology Forum
Standard Sponsored Links

  #2 (permalink)  
Old 01-10-2008, 08:53 PM
danfive's Avatar
Nobel Laureate
Points: 2,293, Level: 30Points: 2,293, Level: 30Points: 2,293, Level: 30
Activity: 100%Activity: 100%Activity: 100%
 

Join Date: Jul 2007
Location: Houston TX
Posts: 507
danfive RSS Feed
Default Re: Problem with extraction of DNA from Trizol

Quote:
Originally Posted by max_rempel View Post
I was able to extract and use RNA from these samples. Now I am trying to extract DNA from the rest of each trizol sample.
The first try gave me very poor quality and low yields of DNA. ?
I had the same thing happen, so I abandoned this method. I also noticed that the RNA from Trizol Extraction contains DNA and therefore needs Dnase I treatment prior to downstream applications.

My advice is to talk with Tech support at Invitrogen.
Digg this Post!Add Post to del.icio.usBookmark Post in TechnoratiFurl this Post!Spurl this Post!Reddit!
Reply With Quote
  #3 (permalink)  
Old 01-11-2008, 02:39 PM
Pipette Filler
Points: 201, Level: 4Points: 201, Level: 4Points: 201, Level: 4
Activity: 0%Activity: 0%Activity: 0%
 

Join Date: Jan 2008
Posts: 4
max_rempel RSS Feed
Default Re: Problem with extraction of DNA from Trizol

I tried another method, it seems to work ok, not sure what is the relatvie yield, but there is at least something.
Remove upper phase. Add 1/3 volume of 70% ethanol.
Load on Qiagen DNAeasy minispin column from Blood and tissue kit.
Continue with washes as described in the kit.
About 50 mg of epidermis gives about 15 mkg of DNA with reasonable , not too clean, quality of DNA by Nanodrop.
I was also given an advice to adjust pH of Trizol phase before extraction. Didn't try yet.
Digg this Post!Add Post to del.icio.usBookmark Post in TechnoratiFurl this Post!Spurl this Post!Reddit!
Reply With Quote
Reply

Tags
dna extraction, trizol

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
Plant Protein Extraction the Hard Way danfive Protein Science 0 04-24-2008 08:53 PM
RNA Kit v.s. Trizol Method c1bl RNA Techniques Forum 5 02-01-2008 07:28 PM
Problems with RNA extraction or QPCR for AI deki Real-Time PCR and Quantitative PCR Forum 2 12-30-2007 07:58 PM
Problems with RNA extraction or QPCR for AI deki Basic Lab Protocols and Techniques 3 07-27-2007 02:59 AM
2D gel running problem cosmis85 Proteomics Forum 2 05-25-2007 06:41 AM


All times are GMT. The time now is 08:04 PM.


Powered by vBulletin® Version 3.7.1
Copyright ©2000 - 2008, Jelsoft Enterprises Ltd.
Copyright 2005-2007 Molecular Station | All Rights Reserved