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  • Maximize the input amounts of RNA where possible.

  • Use 1 hour incubation time for reverse transcription (step 4).

  • Use 48 µl of Purification Solution (PS) at step 16.

  • Perform a qPCR assay to determine the optimal cycle number for library amplification.

See these FAQs for further details:

What do I need for the qPCR Assay?

How do I calculate the Endpoint PCR cycle number from the qPCR assay?

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