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