Strain Protocols

Strain Information
Strain Code: 01XN2
Strain Name: FVB;129S6-Stk11tm1Rdp/Nci
Gene Symbol: Stk11
Protocol Information
Allele: Stk11<tm1Rdp>
Protocol Number: 1
Introductory Comment: This strain carries a functional conditional allele of Lkb1 (official: Stk11) with LoxP sites flanking exons 3 and 6. The genotyping strategy is to detect the upstream LoxP site by flanking it with primers PCRS5 and LKB39. Primers LKB36 and LKB39 produce a wt band, and serve as an internal control. According to the investigator, this primer pair sometimes produces a non-specific 600bp band in -/- mice. Since we expect all mice to be homozygous for this strain, it is not necessary to include the internal control, then.

Primers

PCRS5 : 5'-tct aac aat gcg ctc atc gtc atc ctc ggc-3'
LKB36 : 5'-ggg ctt cc acct ggt gcc agc ctg t-3'
LKB39 : 5'-gag atg ggt acc agg agt tgg ggc t-3'
Diagram showing the primer relataionships for Strain 01XN2.  This strain carries a functional conditional allele of Lkb1 (official: Stk11) with LoxP sites flanking exons 3 and 6.  The genotyping strategy is to detect the upstream LoxP site by flanking it with primers PCRS5 and LKB39.  Primers LKB36 and LKB39 produce a wt band, and serve as an internal control.  According to the investigator, this primer pair sometimes produces a non-specific 600bp band in -/- mice.  Since we expect all mice to be homozygous for this strain, it is not necessary to include the internal control, then.

Product Sizes

Primer Combination Product Size
PCRS5 / LKB39 300 bp    Flox
LKB39 / LKB36 220 bp    WT

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2 2.0 mM
2.5mM dNTP Mix 0.2mM
100 µMPCRS5 0.5 µM
100 µMLKB36 0.5 µM
100 µMLKB39 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
65ºC 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.