CD19Cre Genotyping PCR Protocol

Primers:

CD19.8:            5’ – AAT GTT GTG CTG CCA TGC CTC – 3’
CD19.9:            5’ – GTC TGA AGC ATT CCA CCG GAA – 3’
CD19CreRev:   5’– TTC TTG CGA ACC TCA TCA C – 3’

Reaction Mix for 25ul rxn vol.:

10x PCR bffr       = 2.5ul
50mM MgCl2     = 0.75ul
10mM dNTPs     = 0.5ul
20 uM CD19.8    = 0.25ul
20uM CD19.9     = 0.25ul
20uM CD19CreRev   = 0.25ul
Taq DNA Pol       = 1ul
ddH2O                 = 17.5ul

 Cycling Conditions:

-1x      94 C – 3min
-35x     94 C – 1min
52 C – 1min
72 C – 1min
-1x      72 C – 7min
-1x      4 C – HOLD

WT = ~550b.p.

CRE (CD19null) = ~200b.p.