Strain Protocols

Strain Information
Strain Code: 01XM9
Strain Name: B6;CB-Tg(Fabp7-creLacZ)3Gtm/Nci
Gene Symbol: CreLacZ
Protocol Information
Allele: Tg(Fabp7-creLacZ)3Gtm
Protocol Number: 1
Introductory Comment: This strain contains a Cre recombinase gene under the control of the mouse brain lipid binding protein (BLBP) in order to over-express Cre in neuronal stem cells. Also included is an internal ribosome entry site to allow for co-expression of Cre and LacZ from the same transcript. For this strain, we will simply assay for the presence of the Cre transgene. Primers C001 / C002 amplify the Cre gene, while primers C003 / C004 amplify an endogenous mouse gene, acting as an internal positive control.

Primers

C001 : 5'-acc agc cag cta tca act cg-3'
C002 : 5'-tta cat tgg tcc agc cac c-3'
C003 : 5'-cta ggc cac aga att gaa aga tct-3'
C004 : 5'-gta ggt gga aat tct agc atc atc c-3'
Diagram showing the primer relataionships for Strain 01XM9.  This strain contains a Cre recombinase gene under the control of the mouse brain lipid binding protein (BLBP) in order to over-express Cre in neuronal stem cells.  Also included is an internal ribosome entry site to allow for co-expression of Cre and LacZ from the same transcript.  For this strain, we will simply assay for the presence of the Cre transgene.  Primers C001 / C002 amplify the Cre gene, while primers C003 / C004 amplify an endogenous mouse gene, acting as an internal positive control.

Product Sizes

Primer Combination Product Size
C001 / C002 199 bp    Cre
C003 / C003 300 bp    Internal Control

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2 2.0mM
2.5mM dNTP Mix 0.2mM
100 µMC001 0.5 µM
100 µMC002 0.5 µM
100 µMC003 0.5 µM
100 µMC004 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.