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.


How do you lay out your PCR plates?

How do you lay out your PCR plates? - PCR - Polymerase Chain Reaction Forum

How do you lay out your PCR plates? - 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 07-22-2011, 08:13 PM
Pipette Filler
Points: 4, Level: 1 Points: 4, Level: 1 Points: 4, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jul 2011
Posts: 1
Thanks: 0
Thanked 0 Times in 0 Posts
Default How do you lay out your PCR plates?



When doing qRT-PCR, is it better to run each gene (housekeeping, and one or more experimental genes plus NTCs) on a 96 well plate and maybe only be able to fit six subjects on that plate or is it better to run one gene per plate but fit on all your subjects?

I've heard of people doing it both ways, but it seems more convenient if you run one gene per plate you can use the same housekeeping data over and over again without having to run it in parallel with every new transcript you decide to analyze. I am starting a PCR experiment and I want to make sure I lay my plates out properly.
Reply With Quote
  #2  
Old 08-03-2011, 10:58 PM
Pipette Filler
Points: 799, Level: 15 Points: 799, Level: 15 Points: 799, Level: 15
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Aug 2010
Posts: 20
Thanks: 0
Thanked 7 Times in 6 Posts
Default Re: How do you lay out your PCR plates?

Definitely run one one gene per plate. If you run many genes but fewer samples per plate, you need to to interplate calibration on many plates, which can end up being a nightmare.
Reply With Quote
The Following User Says Thank You to sunglassesatnight For This Useful Post:
admin (08-04-2011)
  #3  
Old 08-20-2011, 05:15 AM
Volunteer
Points: 231, Level: 4 Points: 231, Level: 4 Points: 231, Level: 4
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Aug 2011
Location: Malaysia
Posts: 36
Thanks: 0
Thanked 14 Times in 13 Posts
Default Re: How do you lay out your PCR plates?

I did several plates for 1 gene due to the number of samples and replicates for that experiment including interplate control as calibrator without any nightmare. Main reason is the software i use for analysis of the expression- qBase plus software programme. The software auto-calibrates the variation in different plates. alternatively, you can manually adjust the Ct values of your interplate controls so that they have similar Ct values for each plate.
Reply With Quote
  #4  
Old 08-26-2011, 08:59 PM
Pipette Filler
Points: 799, Level: 15 Points: 799, Level: 15 Points: 799, Level: 15
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Aug 2010
Posts: 20
Thanks: 0
Thanked 7 Times in 6 Posts
Default Re: How do you lay out your PCR plates?

Yes JennYeap you are right. I have also used multiple plates because of large sample size, but without qbase. It's not difficult when you have to calibrate between two or even three plates, but if you have 30 samples on different plates, it quickly turns into a gongshow of interplate calibration madness.
Reply With Quote
Reply

Tags
lay , pcr , plates


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
PAGE gel sticked on both plates Patricia C Arbour-Reily Protocols and Methods Forum 0 12-08-2008 03:41 PM
Simultaneous bagging on 15 plates, N2 and 3 other strains Kathryn Hedges C Elegans Forum 0 11-12-2006 01:49 PM
Problem with -His plates Anna Heidenblut Yeast Forum 0 07-21-2006 03:21 PM
The Classical Vacuum - by Timothy H. Boyer Laurent Physics Forum 8 01-14-2004 03:49 PM
worms on 24-well plates abd elnasser C Elegans Forum 0 07-17-2003 02:23 PM


All times are GMT. The time now is 12:23 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 0.13400 seconds with 16 queries