What is the maximum read lengths and total number of cycles for MiSeq kits?

Illumina platforms and sequencing kits allow for a variety of sequencing applications. To ensure the highest level of quality, Illumina supports reads up to a certain length depending on the sequencing platform and SBS kit version.

It is possible to choose a longer read length during run setup in Local Run Manager (LRM) or the instrument control software. However, when the read length exceeds the supported length, Illumina cannot guarantee the data quality.

The table below lists the maximum supported read lengths for the different MiSeq kits.

Reagent Type

Maximum Read Length

Maximum Index Length

Reagent Kit Size

Maximum Number of Total Cycles

Additional Cycles Needed for Dual Index?

MiSeq v2

51

Not restricted by software

50 cycles

79

Yes (7 cycles)

MiSeq v2 (Including Micro and Nano Kits)

151*

Not restricted by software

300 cycles

329

Yes (7 cycles)

MiSeq v2 (Including Nano Kits)

251

Not restricted by software

500 cycles

529

Yes (7 cycles)

MiSeq v3

151

Not restricted by software

150 cycles

179

Yes (7 cycles)

MiSeq v3

301

Not restricted by software

600 cycles

629

Yes (7 cycles)

\*Note: if performing single read sequencing, can run up to 1x251 bp read length.

For any feedback or questions regarding this article (Illumina Knowledge Article #2595), contact Illumina Technical Support techsupport@illumina.com.

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