12/22/2022
为上机测序提供足够量的SBS试剂是确保测序获得成功的重要因素之一。如果运行过程中试剂量不够，会导致荧光信号强度和测序质量值降低，甚至可能引起测序失败。双端测序在不同测序仪上有不同的操作流程，具体可以参照Indexed Sequencing Overview Guide。在某些仪器上运行双端index测序时，还需要额外七个循环的试剂做第二个index测序前的化学合成反应（仅合成不拍照），此时计算试剂盒所能支持的测序循环数时，一定要考虑到这七个循环也要被包含在总循环数内。
下面表格列举出了多种试剂盒的规格及每种试剂盒最多支持的循环数，请将在某些仪器上运行双端index测序时额外的7个循环考虑在内:
| Reagent Type | Reagent Kit Size | Maximum Number of Cycles | Additional Cycles Needed for Dual Index? | |
|---|---|---|---|---|
| iSeq 100 | i1 (v1 or v2) | 300 | 322 | No |
| MiniSeq | Rapid Kit | 100 | 128 | Yes - 7 cycles |
| High Output 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 Output or Mid Output | 75 | 92 | No |
| 150 | 168 | |||
| 300 | 318 | |||
| HiSeq 1000/1500/2000/2500 | Rapid SBS v2 | 50 | 79 | 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 | |||
| NextSeq 1000/2000 | ||||
| P1 | 100 | 138 | No | |
| 300 | 338 | |||
| 600 | 638 | |||
| P2 | 100 | 138 | No | |
| 200 | 238 | |||
| 300 | 338* | |||
| 600 | 638 | |||
| P3 | 50 | 88 | No | |
| 100 | 138 | |||
| 200 | 238 | |||
| 300 | 327* | |||
| HiSeq 3000/4000 | N/A | 50 | 79 | 7 cycles required for single-read flow cells |
| 150 | 179 | |||
| 300 | 329 | |||
| NovaSeq 6000 | SP v1.0 | 100 | 130 | Yes - 7 cycles |
| 200 | 230 | |||
| 300 | 330 | |||
| 500 | 530 | |||
| S1 and S2 v1.0 | 100 | 130 | ||
| 200 | 230 | |||
| 300 | 330 | |||
| S4 v1.0 | 200 | 230 | ||
| 300 | 330 | |||
| 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 | |||
*The NextSeq 1000/2000 P1和P2试剂盒含有338个cycles 而 P3 v3 试剂盒含有327个cycles.
关于每种测序试剂盒的详细信息请参阅对应的试剂支持页面，不同测序试剂盒支持的最长读长请参阅Maximum read length for Illumina sequencing platforms