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

Real-Time PCR and Quantitative PCR Forum Real-Time PCR and Quantitative PCR Forum


change in CP of included standart with discarded standarts - a standart curve problem

change in CP of included standart with discarded standarts - a standart curve problem - Real-Time PCR and Quantitative PCR Forum

change in CP of included standart with discarded standarts - a standart curve problem - Real-Time PCR and Quantitative PCR Forum


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 12-28-2009, 01:52 PM
Pipette Filler
Points: 54, Level: 1 Points: 54, Level: 1 Points: 54, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Dec 2009
Posts: 1
Thanks: 1
Thanked 0 Times in 0 Posts
Exclamation change in CP of included standart with discarded standarts - a standart curve problem



I am trying to quantify archaea by q-pcr. First I wanted to see how my standarts work . I used 6 standarts I obtained a standart curve at the end of the reaction. I wanted to decrease the error by discarding some of the standarts from calculation. I eliminated two of the standarts. However after I discarded some of the standarts the crossing points of the included ones changed. How could it be possible? I did the same thing before in another q-pcr machine, CP values didn't change. By the way I started to use Light Cycler 480.
Reply With Quote
  #2  
Old 12-28-2009, 03:16 PM
Aga's Avatar
Aga Aga is offline
Post-Doc
Points: 2,239, Level: 30 Points: 2,239, Level: 30 Points: 2,239, Level: 30
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Nov 2008
Posts: 270
Thanks: 25
Thanked 78 Times in 64 Posts
Default Re: change in CP of included standart with discarded standarts - a standart curve pro

The software probably uses dynamic approach and when you eliminated some standards from the curve it calculated everything again. I guess the software dynamically adjusted crossing threshold and so it yielded different Cps.
I'm just guessing off course, but this may be the case. Threshold line is usually default in LC 480 but you can manually set it as you like. Try to look it up in your manual - it should be somewhere where the fit points approach is discussed.
Reply With Quote
The Following User Says Thank You to Aga For This Useful Post:
gozde (01-01-2010)
Reply

Tags
change , curve , discarded , included , problem , standart , standarts


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
Simply put, MOVING DIMENSIONS THEORY is THE NEW MODEL: http://physicsmathforums.com drelliot@gmail.com Physics Forum 0 08-29-2006 06:32 AM
21st Century Dynamics Donald G. Shead Physics Forum 5 08-14-2003 11:43 PM


All times are GMT. The time now is 11:17 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 0.13287 seconds with 16 queries