SyBR green home mix (Duncan Clark)

[Steve Crampton]

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