Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Molecular Biology Techniques
Register Search Today's Posts Mark Forums Read

Molecular Biology Techniques Molecular Biology Forum. Includes forums for common molecular biology techniques.


question about home made sybr green master mix

question about home made sybr green master mix - Molecular Biology Techniques

question about home made sybr green master mix - Molecular Biology Forum. Includes forums for common molecular biology techniques.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 05-27-2011, 05:54 PM
Pipette Filler
Points: 330, Level: 6 Points: 330, Level: 6 Points: 330, Level: 6
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: May 2011
Posts: 4
Thanks: 1
Thanked 0 Times in 0 Posts
Default question about home made sybr green master mix



I tried home made syber green master mix which's recipe was posted by Duncan Clark as below.

makes 10mL store in aliquots in the -20

Stock reagent Volume(10mL) Final
1M Tris pH 8.3 1mL 100mM (UV irradiated)
25mM MgCL2 (Qiagen) 2.4mL 6mM
10mg/mL BSA Omnipure Fraction V 1mL 1mg/mL
10mM dNTP Mix 400μL 400μM
100X SyBR Green 66.67uL 0.66X
dH2O (irradiated DEPC) 5.032 mL

**for use: Add 10μL Qiagen HotStarTaq per 1mL Master Mix



We got pretty good amplification curve and very wired dissociation curve with the home made master mix. The wired dissociation curve is not a curve at all. I could not see where is the baseline, where is the main peak. BTW, all the primers have been tested. I run real-time with ABI syber green master mix on the same plate, all the ABI's wells run perfectly.

Does any one has the same problem? Or any suggestion for this issue?

Thanks a lot.
Reply With Quote
Reply

Tags
green , home , made , master , mix , question , sybr


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
SYBR Green and large amplicons Eli Boritz Real-Time PCR and Quantitative PCR Forum 0 12-22-2010 12:36 AM
Politics And Cannibalism? Introducing The Dourties, Chelsea, Bill, Hillary, Barrack Obama, George Bush, Jr., And All Of Capital Hill! jon_johnfrancisayres@yahoo.com Microbiology Forum 0 10-06-2007 05:59 AM


All times are GMT. The time now is 07:34 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 0.13637 seconds with 16 queries