Pooling recommendations for the AmpliSeq for Illumina Myeloid Panel

The AmpliSeq for Illumina Myeloid Panel creates libraries from DNA and RNA. Refer to the table below for instrument specific pooling recommendations.

Instrument	Max DNA only	Max RNA only	Max DNA/RNA	Pooling volume ratio
MiniSeq MO	4	32	3 DNA + 3 RNA	8 DNA : 1 RNA
MiniSeq HO	12	96	11 DNA + 11 RNA
MiSeq v2	7	60	6 DNA + 6 RNA
MiSeq v3	12	96	11 DNA + 11 RNA

What is the recommended coverage?

• Illumina recommends 1000x minimum coverage, and 6000x mean coverage.

  • This equates to approximately 2 million reads per DNA sample and 0.25 million reads per RNA sample.

  • As an example: a MiSeq v3 run provides ~25M reads. - Combine DNA:RNA at 8:1 (8 µl of DNA final library to 1 µl of RNA final library). - We target 2M reads per DNA library + 0.25M reads per RNA library = 2.25M reads / combined sample. - 25 million reads per MiSeq v3 run / 2.25 million = 11 paired samples per MiSeq v3 run.

See the AmpliSeq for Illumina Myeloid Panel Compatible Products page for more information.

For any feedback or questions regarding this article (Illumina Knowledge Article #1047), contact Illumina Technical Support [e-mail].