Strain Protocols

Strain Information
Strain Code: 01P05
Strain Name: C3;B6-Tg(CTSG-PML/RARa)135Ley/Nci
Gene Symbol: PML/RARA
Protocol Information
Allele: Tg(CTSG-PML/RARA)135Ley
Protocol Number: 1
Introductory Comment: This strain carries a chimeric promyelocytic leukemia-retinoic acid receptor alpha transgene (PML-RARa) under the of the human cathepsin G gene. The strategy of this genotyping protocol is to detect the human cathepsin G gene with primers P005 and P006. Primers P007 and P008 are included as an internal PCR control and amplify the endogenous mouse colony stimulating factor 3 receptor gene (csf3r).

Primers

P005 : 5'-GGc cTG Acc TcA Tcc cAT AG-3'
P007 : 5'-cTG cTc TTG AcA TAA Gcc TG-3'
P006 : 5'-Gcc cTT TTc ccc ATc cTA GG-3'
P008 : 5'-TAG cAG cAc TcT TcA ATG Gc-3'

Product Sizes

Primer Combination Product Size
P005 / P006 520 bp    CTSG Promoter
P007 / P008 360 bp    Internal Control

Master Mix

Reagent Final Concentration
10X Buffer 1X
25mM MgCl2 2.0 mM
2.5mM dNTP Mix 0.2 mM
DMSO 1%
100 µMP005 0.25 µM
100 µMP007 0.25 µM
100 µMP006 0.25 µM
100 µMP008 0.25 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 1:00
60ºC 2:00 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.