12/10/2022
Maximum supported read length
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; refer to the How many cycles of SBS chemistry are in my kit? support bulletin. 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 maintaining high data quality.
Table 1. Maximum supported read length for sequencing platforms and SBS reagent kits.
Sequencing Platform | SBS Kit Version | Maximum Read Length |
---|---|---|
iSeq 100 | v1 | 2 x 151bp |
v2 | 2 x 151bp | |
MiniSeq | MO* | 2 x 151bp |
HO* | 2 x 151bp | |
MiSeq | v2 | 2 x 251bp |
v3 | 2 x 301bp | |
NextSeq 500/550 | MO* | 2 x 151bp |
HO* | 2 x 151bp | |
NextSeq 1000/2000 | P1, P2 | 2 x 301bp |
P3 | 2 x 151bp | |
HiSeq 1000/1500/2000/2500 | HO* v3 | 2 x 101bp |
HO* v4 | 2 x 126bp | |
RR** v4 | 2 x 251bp | |
HiSeq 3000/4000 | N/A | 2 x 151bp |
HiSeq X | N/A | 2 x 151bp |
NovaSeq 6000 | SP | 2 x 251bp |
S1, S2, S4 | 2 x 151bp |
* MO: Mid-output / HO: High-output
** Rapid Run
Maximum read length for index reads
Table 2. Maximum individual index length for sequencing platforms.
Sequencing Platform | Single or Dual Index (Maximum Length) |
---|---|
iSeq 100 | Not restricted by software |
MiniSeq v1.X | 20 bp |
MiniSeq v2.X | 20 bp |
MiSeq < v4 | Not restricted by software |
MiSeq ≥ v4 | 100 bp** |
NextSeq 500/550 v2 NCS | 20 bp |
NextSeq 500/550 v4 NCS | 20 bp |
NextSeq 1000/2000* | Not restricted by software |
HiSeq 2500 | Not restricted by software |
HiSeq 3000/4000 | Not restricted by software |
HiSeq X | 6 bp Single Index / 2 x 8 bp Dual Index |
NovaSeq 6000 < v1.6.0 | 20 bp |
NovaSeq 6000 ≥ v1.6.0 | Not restricted by software |
*When setting up runs in Cloud Mode, index length is limited to 10 bp.
**For index length between 20-100 bp, use manual mode as LRM v3 is limited to 20 bp