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How many cycles of SBS chemistry are in my kit?

Making sure that sufficient volumes of SBS reagents are loaded onto the sequencing instrument is crucial for a successful run. Running out of reagents during a run impacts intensity and quality, and can result in a run failure. Dual-index sequencing, depending on the instrument, follows different workflows, as shown in the Indexed Sequencing Overview Guide. On certain instruments, dual-index runs need an additional seven cycles of reagents for the chemistry-only cycles at the beginning of the second Index Read. The seven chemistry-only cycles are required for the workflows in which the second Index Read sequencing occurs after the forward template anneals to the grafted P5 primer on the surface of the flow cell. These seven cycles must be considered when calculating the number of cycles supported by the kit.
The following table outlines available reagent kit sizes and the maximum number of cycles each kit can perform, taking the additional seven cycles into consideration:
Instrument
Reag. Kit Type
Kit Size
Max No. of Cycles
Dark Cycles Needed for Dual Index?
iSeq
v2
300
322
No
MiniSeq
Rapid Kit
100
128
Yes - 7 Cycles
High or Mid Output
75
92
No
150
168
300
318
MiSeq
V2 (Including Micro and Nano Kits)
50
79
Yes - 7 Cycles
300
329
500
529
V3
150
179
600
629
NextSeq 500/550
High or Mid Output
75
92
No
150
168
300
318
NextSeq 1000/2000
P1
100
138
No
600
638
P2
100
138
200
238
300
338*
600
638
P3
50
88
100
138
200
238
300
327*
HiSeq 1000/1500/2000/2500
Rapid SBS V2
50
79
Yes - 7 Cycles required for paired-end flow cells
200
229
500
529
TruSeq SBS V3
50
58
200
209
HiSeq SBS V4
50
79
250
279
HiSeq 3000 / 4000 / X
N/A
50
79
Yes - 7 Cycles required for single-read flow cells
150
179
300
329
NovaSeq 6000
SP v1.5
100
138
No
200
238
300
338
500
538
S1 and S2 v1.5
100
138
200
238
300
338
S4 v1.5
35
72
200
238
300
338
NovaSeq X / X Plus
B10
100
138
No
200
238
300
338
*The NextSeq 1000/2000 P2 v3 kit has 338 total cycles while the P3 v3 kit has 327 total cycles.
For more information about each SBS reagent kit, including how to prepare, combine, and split SBS reagents when possible, visit the support page for each kit; also refer to the bulletin Maximum read length for Illumina sequencing platforms for maintaining high data quality.
For any feedback or questions regarding this article (Illumina Knowledge Article #7002), contact Illumina Technical Support [email protected].
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