RNA amplification - (Feb/19/2008 )

Hi all


can anyone advise me on how to amplify the total RNA of my samples. I get very small amounts of sample giving me RNA sometimes as low as only 100 ngs. Need lot more of it to screen various genes via Real time PCR.

Many thanks
Rits

---

Please describe the way you are doing it.

---

I am currently extracting RNA from human white blood cells using RNeasy micro kit. Since the blood sample volumes are very limiting (2-5 mL) only the RNA yield is very low.

Cheers

---

hi
i think best method for you will be to use two step Real time PCR
use some sensitive Reverse transcriptase kit like Sensiscript RT kit from qiagen with random nanomers as primer for reverse transcription so that you can reverse transcribe whole transcriptome of the cells (though they recommend using gene specific primers but as your RNA is very less in amount you have to compromise on it).

Best of luck

---