Mycoplasma test from Blasi lab

• 1mL conditioned medium (at least 24hrs in culture)

• spin 5' at 1200rpm

• keep supernatant

• spin supernatant 15' 13000 rpm at 4°C

• resuspend pellet in 20µL H20

• boil sample 10'

• do PCR with 2µL of sample

PCR program:

5' 94°C

[ 30'' 94°C, 30'' 56°C, 45'' 72°C ] x30 cycles

10' 72°C

Primers:

Myc FW: ACTCCTACGGGAGGCAGCAGT

Myc RW: TGCACCATCTGTCACTCTGTTAACCTC

• Amplified band size is around 720 bp