What recommendations do you have for targeting rare, or low-abundance transcripts?

• 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 endpoint cycle numbers?