Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Epigenetics Forum: DNA Methylation, Histone and Chromatin Study
Register 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


bisulfite + taqman- hellpppp...

bisulfite + taqman- hellpppp... - Epigenetics Forum: DNA Methylation, Histone and Chromatin Study

bisulfite + taqman- hellpppp... - An Epigenetics Forum Dealing with DNA Methylation, the study of histones and chromatin


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 11-06-2008, 06:37 PM
Pipette Filler
Points: 226, Level: 4 Points: 226, Level: 4 Points: 226, Level: 4
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Nov 2008
Posts: 8
Thanks: 0
Thanked 0 Times in 0 Posts
Default bisulfite + taqman- hellpppp...



i'm trying to perform bisulfite conversion and real time PCR afterwards in order to find out methylation of BRCA1 gene.
im using cell lines of lung and breast cancer and convert only 1 ug of extracted DNA of each cell line, then im doing real time PCR using primers and prob of ALU gene (for start, just to see if i have enough DNA) . im using the taqman for the real time...
results of the real time: 2 times i got nothing and 2 other times i got amplification but the CT was too high...

can anybody help? pls if you have any experience with this I would be grateful...Im pretty desperate...

thanx! (sorry if I have any english mistakes...)
Reply With Quote
  #2  
Old 11-14-2008, 10:29 PM
Pipette Filler
Points: 1,547, Level: 23 Points: 1,547, Level: 23 Points: 1,547, Level: 23
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Feb 2008
Posts: 13
Thanks: 0
Thanked 1 Time in 1 Post
Default Re: bisulfite + taqman- hellpppp...

Hi, I did few experiments with bisulfite for methylation studies but only using regular PCR and cloning or pyrosequencing(by a compagny). Did you try regular PCR? 1ug of genomic DNA is a good amount to work with. Are the primers you are using have been used before?
Reply With Quote
  #3  
Old 11-15-2008, 06:13 PM
Pipette Filler
Points: 226, Level: 4 Points: 226, Level: 4 Points: 226, Level: 4
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Nov 2008
Posts: 8
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: bisulfite + taqman- hellpppp...

yep... the primers and prob are working for sure... I dont know what I am doing wrong.

can you give more details about the regular PCR that you are doing? how it helps you for detecting the % methylation?
Reply With Quote
Reply

Tags
bisulfite , hellpppp , taqman


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
which kit is 2 b used for bisulfite conversion?? METHY Epigenetics Forum: DNA Methylation, Histone and Chromatin Study 1 06-24-2010 07:54 AM
bisulfite primers labpandit Epigenetics Forum: DNA Methylation, Histone and Chromatin Study 0 03-16-2010 11:48 AM
Help with PCR for bisulfite converted DNA!!! punkyneen Epigenetics Forum: DNA Methylation, Histone and Chromatin Study 2 03-11-2010 06:58 AM
real time PCR + taqman program shir Real-Time PCR and Quantitative PCR Forum 2 07-21-2009 10:49 AM
bisulfite treatment for methylation-sensitive PCR biomol@inca.gov.br Protocols and Methods Forum 0 11-23-2004 07:45 PM


All times are GMT. The time now is 06:49 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 0.12282 seconds with 16 queries