IMB205

ImProm-II™ Reverse Transcription System

Experimental Reaction

Part I

X μl              Experimental RNA (up to 1μg/reaction)

X μl              Primer (10~20pmol)

X μl              Nuclease-Free Water (final reaction volume to 5 μl)

at 70℃ for 5 minutes and chilled on ice (at least 5 minutes).

Part II

4 μl              5X Reaction Buffer

X μl              MgCl2 (final concertration 1.5~8.0mM)

1 μl               dNTP (final concertration 0.5mM)

1 μl               RNase inhibitor (optional)

X μl              Nuclease-Free Water (final reaction volume to 15 μl)

1 μl               Reverse Transcriptase

chilled on ice

Part I + Part II

at 25℃ for 5 minutes

at 42℃ for up to 1 hour (optimized between 37℃ and 55℃).

at 70℃ for 15 minutes (Inactivate Reverse Transcriptase)

Download this file : http://hyuan.imb.sinica.edu.tw/protocol/RT.doc