How many cycles of SBS chemistry are in my kit?

04/02/20


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:

Reagent Type Reagent Kit Size Maximum Number of Cycles Additional Cycles Needed for Dual Index?
HiSeq - TruSeq SBS v3 50 58 7 cycles required for paired-end flow cells
200 209
HiSeq - HiSeq SBS v4 50 79
250 279
HiSeq - Rapid SBS v2 50 79
200 229
500 529
MiSeq v2 (including Micro and Nano kits) 50 79 Yes - 7 cycles
300 329
500 529
MiSeq v3 150 179
600 629
NovaSeq 6000 SP 100 130
200 230
300 330
500 530
NovaSeq 6000 S1 and S2 100 130
200 230
300 330
NovaSeq 6000 S4 200 230
300 330
HiSeq 3000/4000 SBS 50 79 7 cycles required for single-read flow cells
150 179
300 329
iSeq 100 i1 300 322 No
MiniSeq High Output or Mid Output 75 92
150 168
300 318
NextSeq 500/550 High Output or Mid Output 75 92
150 168
300 318

 

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.