Requirements & Compatibility

The NeoPrep System normalizes libraries to 10nM.

  • Dilute the normalized library to the predenature concentration specified (eg, 4 nM for NextSeq).
  • Denature and dilute the library to an intermediate concentration specified (eg, 40 pM for NextSeq). Depending on the application, pooling after denaturation and dilution can increase library complexity due to the number of unique molecules present in the final mixture when each library is denatured separately in a larger volume.
  • Load the denatured samples at the specified load concentration (eg, 2–3 pM for NextSeq)

TruSeq Stranded mRNA Library Prep for NeoPrep libraries result in the following cluster densities:

Instrument

Insert Size

Predenature concentration (nM)

Denatured/diluted intermediate concentration (pM) Pooled denatured samples pM load

Expected Cluster Density (mm2)

MiSeq

 

200

10

50 10

1200

NextSeq

200

4

40 2–3

~200

HiSeq 2500 High Output v3

 

200

2

20 6–10

700–1000

HiSeq 2500 High Output v4

 

200

2

20 12–15

1250

HiSeq 2500 Rapid Run v1

 

200

2

20 6

600–1000

For sequencing instrument compatibility information, see the Illumina Version Compatibility Reference.