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. On certain instruments, dual-index runs need an additional 7 cycles of reagents for the chemistry-only cycles at the beginning of the second Index Read. These 7 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:
|Reagent Type||Reagent Kit Size||Maximum Number of Cycles||Single-Indexed Example|
|HiSeq - TruSeq SBS v3||50||58||7 cycles required for paired-end flow cells|
|HiSeq - HiSeq SBS v4||50||75|
|HiSeq - Rapid SBS v2||50||75|
|MiSeq v2 (including Micro and Nano kits)||50||75||Yes - 7 cycles|
|NovaSeq 6000 S1 and S2||100||130|
|NovaSeq 6000 S4||300||330|
|HiSeq 3000/4000 SBS||50||75||7 cycles required for single-read flow cells|
|iSeq 100 i1||300||318||No|
|MiniSeq High Output or Mid Output||75||92|
|NextSeq 500/550 High Output or Mid Output v2 and v2.5||75||92|
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.