Strain Protocols

Strain Information
Strain Code: 01XN5
Strain Name: B6.129S6-Tgfbr2tm1Hlm/Nci
Gene Symbol: Tgfbr2
Protocol Information
Allele: Tgfbr2<tm1Hlm>
Protocol Number: 1
Introductory Comment: This strain is a conditional allele in which Exon 2 of the TGF- type II receptor (Tgfbr2) gene was flanked by a LoxP site. Therefore, the genotyping strategy is to detect this LoxP site with primers that flank it. Note: The Neo gene that was part of the targeting cassette has been recombined out in this allele.

Primers

T004 : 5'-taa aca agg tcc gga gcc ca-3'
T005 : 5'-act tct gca aga ggt ccc ct-3'
Diagram showing the primer relataionships for Strain 01XN5.  This strain is a conditional allele in which Exon 2 of the TGF-  type II receptor (Tgfbr2) gene was flanked by a LoxP site.  Therefore, the genotyping strategy is to detect this LoxP site with primers that flank it.

Note: The Neo gene that was part of the targeting cassette has been recombined out in this allele.

Product Sizes

Primer Combination Product Size
T004 / T005 540 bp    Mutant
T004 / T005 420 bp    wt

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2 2.0 mM
2.5mM dNTP Mix 0.2 mM
100 µMT004 0.5 µM
100 µMT005 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.