Strain Protocols

Strain Information
Strain Code: 01XL5
Strain Name: B6.FVB-Tg(Pdx1-cre)6Tuv/Nci
Gene Symbol: Cre
Protocol Information
Allele: Tg(Ipf1-cre)1Tuv
Protocol Number: 1
Introductory Comment: This strain contains the Pxd-1 promoter element controlling expression of Cre recombinase in the pancreas. The strategy of this genotyping method is to anchor the sense primer in the Pdx promoter, and the antisense primer in the Cre gene. This will produce a more specific PCR product that just simply assaying for Cre with generic Cre primers.

Primers

Pdx-F : 5'-ctg gac tac atc ttg agt tgc-3'
Pdx-R : 5'-ggt gta cgg tca gta aat ttg-3'
Diagram showing the primer relataionships for Strain 01XL5.  This strain contains the Pxd-1 promoter element controlling expression of Cre recombinase in the pancreas.  The strategy of this genotyping method is to anchor the sense primer in the Pdx promoter, and the antisense primer in the Cre gene.  This will produce a more specific PCR product that just simply assaying for Cre with generic Cre primers.

Product Sizes

Primer Combination Product Size
Pdx-F / Pdx-R 650 bp    Transgene

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2 2.0 mM
2.5mM dNTP Mix 0.2 mM
100 µMPdx-F 0.5 µM
100 µMPdx-R 0.5 µM
5U/┬Ál Taq Pol 0.025 U/µl
Use 50 - 100 ng of DNA per reaction

Cycling Conditions

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