Strain Protocols

Strain Information
Strain Code: 01XE7
Strain Name: B6.Cg-Tg(Vil-cre)20Sy/Nci
Gene Symbol: Cre
Protocol Information
Allele: Tg(Vil-Cre)20Syr
Protocol Number: 2
Introductory Comment: This strain contains a Cre recombinase gene linked to the murine villin (Vil) promoter, resulting in selective expression of Cre in the intestinal epithelium. The genotying strategy is to detect this transgene with primer V001, which is specific for the Vil promoter, and V002, which is specific for the Cre recombinase gene.

Primers

V001 : 5'-caa gcc tgg ctc gac ggc c-3'
V002 : 5'-cgc gaa cat ctt cag gtt ct-3'
Diagram showing the primer relataionships for Strain 01XE7.  This strain contains a Cre recombinase gene linked to the murine villin (Vil) promoter, resulting in selective expression of Cre in the intestinal epithelium.  The genotying strategy is to detect this transgene with primer V001, which is specific for the Vil promoter, and V002, which is specific for the Cre recombinase gene.

Product Sizes

Primer Combination Product Size
V001 / V002 290 bp    Vil-Cre Transgene

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2 2.0 mM
2.5mM dNTP Mix 0.2 mM
100 µMV001 0.5 µM
100 µMV002 0.5 µM
5U/┬Ál Taq Pol 0.015 U/µl
Use 50 - 100 ng of DNA per reaction

Cycling Conditions

(Time in minutes:seconds)
Temperature Time Label
94ºC 3:00
0ºC
94ºC 0:30
60ºC 0:30 35 Cycles
72ºC 0:30
0ºC
72ºC 3:00
4ºC HOLD

Disclaimer: PCR conditions were optimized using QIAGEN-purified mouse tail DNA and a PTC-200 thermocycler (MJ Research). Optimal conditions may vary with DNA quality or equipment used.