Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > General Forum > Chit Chat
Register Search Today's Posts Mark Forums Read


pcr

pcr - Chit Chat

pcr -


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 04-01-2012, 05:52 PM
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 pcr



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
pcr


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


All times are GMT. The time now is 09:21 AM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2013, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.14861 seconds with 15 queries