Protocols

qRT-PCR

1. pick up a colony and inoculate in LB broth containing 100µg/mL ampicillin
2. incubate at 37 degree over night
3. dillute the over night culture (1:50) in LB broth containing 100µg/mL ampicillin
4. grow to OD600=0.6 at 37 degree
5. centrifuge 1mL culture at 15,000rpm for 1min
6. remove the supernatant
7. resuspend with TE buffer containing lysozyme(0.4mg/mL)
8. incubate at room temperature for 5min
9. isolate total RNA from the lysate with RNeasy® Mini
10. synthesize cDNA from 1µg total RNA with PrimeScript® 1st strand cDNA Synthesis Kit
11. quantify the cDNA of the target gene or 16S rRNA with Power SYBR® Green PCR Master Mix