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.
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