Master GREEN 200 Manual de usuario Pagina 28

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3-4 Reverse Transcription
Performing RT
Reactions
The procedure for generating cDNA using the TaqMan Reverse
Transcription Reagents is described below.
CHEMICAL HAZARD. TaqMan Reverse Transcription
Reagents may cause eye and skin irritation. They may cause discomfort if
swallowed or inhaled. Always use adequate ventilation such as that provided by
a fume hood. Please read the MSDS, and follow the handling instructions. Wear
appropriate protective eyewear, clothing, and gloves.
To perform RT reactions:
Step Action
1 Prepare the RT Reaction Mix by combining all the nonenzymatic
components.
2 Vortex briefly.
3 Add the enzymatic components (e.g., MultiScribe Reverse
Transcriptase, RNase Inhibitor) and the RNA.
4 Mix the components by inverting the microcentrifuge tube.
5 Transfer the contents to a MicroAmp
®
Optical Tube or multiple
wells of a MicroAmp
®
Optical 96-Well Reaction Plate.
6 Cap the plate/tubes with MicroAmp
®
Optical Caps.
Note Alternatively, you may seal the plate with a MicroAmp
Optical Adhesive Cover. However, do not use the cover with
MicroAmp Optical Tubes.
7 Centrifuge the plate/tubes briefly to remove air bubbles and collect
the liquid at the bottom of the tube.
8 Transfer the plates to the thermal cycler block.
9 Perform RT.
10 Remove the 96-well reaction plate after thermal cycling is complete.
CAUTION
!
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