First Strand cDNA Synthesis (AB Kit): Difference between revisions

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adjusted quantity of RNA added
 
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* Add 10 uL Master mix per well into tube or plate
* Add 10 uL Master mix per well into tube or plate
* Add 0.1-10 ug RNA in a final volume of 10 uL to each well
* Add 10 ug RNA in a final volume of 20 uL to each well. Since results will be normalized in the end, standardizing RNA quantities at this step is unnecessary. Total volume can be adjusted down if RNA is in short supply.
* Run the PCR Program '''AB Reaction Protocol'''.  Takes ~2h:
* Run the PCR Program '''AB Reaction Protocol'''.  Takes ~2h:
** 10 min at 25C
** 10 min at 25C
** 120 min at 37C
** 120 min at 37C
** 5 min at 85C
** 5 min at 85C