Real-time reverse transcription (RT)-PCR

Real-time reverse transcription (RT)-PCR
 

Step-by-step procedure:

A. RNA isolation:

At indicated time points, cells were lysed with TRIzol (500 uL/1.2x106 cells). Total RNA was isolated following the TRIzol reagent (Invitrogen) manufacturer’s instructions.

B. cDNA synthesis:

Prepare the master mix as per detail are given below (per sample)

Note:

     Please follow the PCR cycle as shown in the picture below.

     Dilute the PCR product 10X with ultrapure distilled water.

C. Real-time PCR:

Prepare the real-time PCR master mix as per detail are given below (per sample)

Note:

     Please see the primer sequences details in the respective published paper.

     Please follow the real-time PCR cycle as shown in the picture below.