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.


DNA template and primers concentration

DNA template and primers concentration - PCR - Polymerase Chain Reaction Forum

DNA template and primers concentration - 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 03-14-2012, 12:28 AM
Pipette Filler
Points: 91, Level: 1 Points: 91, Level: 1 Points: 91, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Mar 2012
Location: London
Posts: 3
Thanks: 0
Thanked 0 Times in 0 Posts
Default DNA template and primers concentration



Hi all,


I am struggling in doing PCR , it sounds thai I have some problem . the big problem I have been facing is to calculate the the DNA and primers concentration after measuring it by nanodrop.
for example: wHAT IS written in he bottles as follows:

I have DAN
18.6NM
Mw:21,622

T7 -forward primers:
120.3nm , M w:12.754.3. I have made 100um stock solution by multiple 18.6 *10 += 186 water added

REVERS PRIMERS:
155.6NM M w:5.852.8
I have made 100um stock solution by multiple155.6 *10 and then 1556 water added . I uasually take 10 ul from the stock to add to the 1ML PCR reaction to make 1 um final concentration. however, I did not got any good resuilt yet .
my supervisor told me that I have to make 1fetomolar concentartion from DNA and then use the nanomolar concentration from the both primers ( iam not sure 1 or 10 nanomolar)
how to make this ratio :

is that kind of diluation if so how can i dilute it since i got very unmeasurable number when i use the formula cv=c1v1

what is the right and accurate steps should follow when measuring DNA concentration by nanodro and getting (2.3 ng/ul ) to make the unit nanomolar nMor fetomolar fM

any one can help.

THANKS
Reply With Quote
Reply

Tags
concentration , dna , primers , template


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
Why is one species DNA favored in mixed sample PCR using universal primers brendan101 PCR - Polymerase Chain Reaction Forum 3 09-23-2011 11:28 AM
PCR with DGGE primers fails with soil DNA Nenozabaloy PCR - Polymerase Chain Reaction Forum 0 03-22-2011 03:08 PM
No primers visible on the end of a agarose gel ese PCR - Polymerase Chain Reaction Forum 4 02-13-2010 04:20 PM
hygromycin primers Analia Alet Protocols and Methods Forum 0 10-08-2007 09:24 PM
optimizing overlap PCR, ligation-free cloning Maximilian Haeussler Protocols and Methods Forum 0 05-29-2007 01:56 PM


All times are GMT. The time now is 09:51 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 0.11787 seconds with 16 queries