Alport (Col4A3) Mice Genotyping

 

For Col4A3 wild type mice

 

Genomic DNA                    1 μL

2x PCR Mix                         10 μL

Oligo #824                          1 μL

Oligo #825                          1 μL

Water                                   7 μL

 

Total                                      20 μL

 

94°C                          3 minutes

35 cycles:

94°C                          30 seconds

62°C                          1 minutes

72°C                          2 minutes

 

72°C                          7 minutes

4°C                             

 

Wild Type band: ~810 bp:

 

 

For Col4A3 Mut (KO) mice

 

Genomic DNA                    1 μL

2x PCR Mix                         10 μL

Oligo #826                          1 μL

Oligo #827                          1 μL

Water                                   7 μL

 

Total                                      20 μL

                                   

94°C                          1 min 30 seconds

35 cycles:

94°C                          30 seconds

65°C                          1 min 30 seconds

72°C                          30 seconds

 

72°C                          5 minutes

4°C                             

 

Mutant band: ~280 bp
Disregard non-specific band at ~650 bp