Strain Protocols

Strain Information
Strain Code: 01XA6
Strain Name: STOCK Tgfb1tm1Doe Rag2tm1Fwa/Nci
Gene Symbol: Tgfb1
Protocol Information
Allele: Tgfb1<tm1Doe>
Protocol Number: 1
Introductory Comment:

Primers

T001 : 5'-GAG AAG AAc TGc TGT GTG cG-3'
T003 : 5'-Gcc GAG AAA GTA Tcc ATc AT-3'
T002 : 5'-GTG Tcc AGG cTc cAA ATA TAG G-3'

Product Sizes

Primer Combination Product Size
T001 / T002 180 bp    Wildtype
T002 / T003 600 bp    Knockout

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2 2.0 mM
2.5mM dNTP Mix 0.2 mM
100 µMT001 0.33 µM
100 µMT003 0.33 µM
100 µMT002 0.33 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 1:00
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.