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SyBR green home mix (Duncan Clark)

SyBR green home mix (Duncan Clark) - Protocols and Methods Forum

SyBR green home mix (Duncan Clark) - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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  #1  
Old 02-15-2008, 06:49 PM
Steve Crampton
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Default SyBR green home mix (Duncan Clark)



We use our own QPCR MM that includes the SyBR green from Molecular
probes. This home made mix is much cheaper than premade mixes from
most companies. the reference for the mix is below the recipe. Of
course, you still have to get the Hot start taq (we use Qiagens).
Steve
Quantitative PCR Master Mix (2X)

makes 10mL store in aliquots in the -20

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


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

Rxn
10μL MM
5μL dH2O
2μL upper primer
2μL lower primer
1μL template
20μL


Reference for MM
Phosphoinositide 3-kinase and Bruton's tyrosine kinase regulate
overlapping sets of genes in B lymphocytes.
Fruman DA, Ferl GZ, An SS, Donahue AC, Satterthwaite AB, Witte ON
Proc Natl Acad Sci U S A. 2002 Jan 8;99(1):359-64
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Old 11-06-2012, 09:13 PM
Pipette Filler
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Default Re: SyBR green home mix (Duncan Clark)

Hi,

I'm interested in seeing whether anyone has tried the SyBr green home mix in this post. We've being using IQ SYBR GRN SUPERMIX from Bio-Rad and it's pricy. Also, in the recipe the concentration of SyBR Green is 100x, I wonder what does this mean? Can you buy SYBR Green I from Molecular Probes that says 100x in concentration?

Thanks,
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