Strain Protocols

Strain Information
Strain Code: 01XBF
Strain Name: B6;C3-TgTn(sb-T2/Onc2)6113Njen/Nci
Gene Symbol: T2/Onc2
Protocol Information
Allele: TgTn(sb-T2/Onc2)6113Njen
Protocol Number: 1
Introductory Comment: This strain carries a modified version of the Sleeping Beauty (SB) transposon, which contains a longer Engrailed-2 splice acceptor (En2-SA) element, a bi-directional polyadenylation signal (pA), and a murine stem cell virus long terminal repeat element (MSCV LTR). The genotyping strategy for this strain is to detect the SB transposase binding site in the inverted terminal repeat end with primers T2Onc2-F and T2Onc2-R.

Primers

T2O2F : 5'-cag ttg aag tcg gaa gtt ta-3'
T2O2R : 5'-gga att gtg ata cag tga at-3'
Diagram showing the primer relataionships for Strain 01XBF.  This strain carries a modified version of the Sleeping Beauty (SB) transposon, which contains a longer Engrailed-2 splice acceptor (En2-SA) element, a bi-directional polyadenylation signal (pA), and a murine stem cell virus long terminal repeat element (MSCV LTR).  The genotyping strategy for this strain is to detect the SB transposase binding site in the inverted terminal repeat end with primers T2Onc2-F and T2Onc2-R.

Product Sizes

Primer Combination Product Size
T2O2F / T2O2R 192 bp    Transgene

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2 2.0mM
2.5mM dNTP Mix 0.2mM
100 µMT2O2F 0.5 µM
100 µMT2O2R 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.