TaqMan RT

DNA Free

1. In 26ul of sample, add 3ul of 10X Dnase I buffer and 1ul rDNase I
2. Incubate at 37°C for 20 min.
3. Add 3ul DNase Inactivation Reagent
4. Incubate 2 min at room temp with occasional mixing
5. Spin at 13,000 rpm for 1.5 min at room temperature.
6. Then transfer supernatant to a new tube.

TaqMan RT

1. Calculation the RNA we need in 8ul mix. O.D Conc.

2. Master Mix

3. Mix the 8ul mix with the 12.4ul master mix together into little strip tube.

(The total volume of the reaction is 20ul)

4. Run the reaction with rt-taqman program.

(25°C for 10min => 48°C for 60min => 95°C for 5min => 4°C)

TaqMan-PCR

1. Mix the above solution into the little strip tube.
2. Run the reaction with tap-pcr program. (40 cycle, 50°C for 2min => 95°C for 15min => 95°C for 15sec => 61°C for 1 min => 25°C for 60 min)
3. Stop the reaction at 25°C
4. Run in 2% gel and take picture.