Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > PCR - Polymerase Chain Reaction Forum
Register Search Today's Posts Mark Forums Read

PCR - Polymerase Chain Reaction Forum PCR - Polymerase Chain Reaction Forum. Discuss and ask questions about PCR troubleshooting, PCR protocols and methods, PCR products, and PCR theory.


Questions on RT-qPCR Experimental Design

Questions on RT-qPCR Experimental Design - PCR - Polymerase Chain Reaction Forum

Questions on RT-qPCR Experimental Design - PCR - Polymerase Chain Reaction Forum. Discuss and ask questions about PCR troubleshooting, PCR protocols and methods, PCR products, and PCR theory.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 12-03-2010, 04:30 PM
Pipette Filler
Points: 787, Level: 15 Points: 787, Level: 15 Points: 787, Level: 15
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Dec 2009
Posts: 1
Thanks: 0
Thanked 0 Times in 0 Posts
Default Questions on RT-qPCR Experimental Design



Hello everyone,

I am very new to RT-qPCR.
I am performing RNAi experiment as described below.

I have silenced one gene (gene A) with four different siRNAs. I performed triplicate experiment for each siRNA.
I used GAPDH as a positive control to check for siRNA delivery efficiency.
I performed triplicate experiment for GAPDH too.
I used scrambled siRNA as a negative control. I performed triplicate experiment for scrambled siRNA too.
Please I need help on the way I should arrange the RT-qPCR for:

1. Each 3 samples of the 4 siRNAs = 12 samples
2. The 3 samples of the GAPDH siRNA = 3 samples
3. the 3 samples of the scrambled siRNA = 3 samples
4. Non-template control (NTC) = ? samples

I was informed that I should use another house keeping gene such as ACTB that was not silenced to serve as a control
for the Gene A and GAPDH which I have silenced.
How do I peeform QC for the transfection reagents I used?
I would be happy to receive help from everyone.
I would be happy to received used protocols that are same or similar to what I want to do.

Thank you very much,

Dr. Abubakar
Reply With Quote
Reply

Tags
design , experimental , questions , rtqpcr


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
qPCR NEWS May 2009 - focus on RNA integrity Editor www.Gene-Quantification.info Botany Forum 0 05-28-2009 11:01 AM
qPCR NEWS May 2009 - focus on RNA integrity Editor www.Gene-Quantification.info Protocols and Methods Forum 0 05-28-2009 10:58 AM
qPCR NEWS January 2009 - update in available real-time PCR cycler Editor www.Gene-Quantification.info Protocols and Methods Forum 0 02-19-2009 11:04 AM
qPCR NEWSLETTER - September 2007 - qPCR 2007 talks online Editor www.Gene-Quantification.info Protocols and Methods Forum 0 09-25-2007 12:34 PM
qPCR 2007 - CALL for scientific contributions Editor www.Gene-Quantification.info Protocols and Methods Forum 0 01-22-2007 02:24 PM


All times are GMT. The time now is 12:53 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 0.11815 seconds with 16 queries