我所购买的SBS试剂盒最多能够满足多少个循环的测序?

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 TypeReagent Kit SizeMaximum Number of CyclesAdditional Cycles Needed for Dual Index?

iSeq 100

i1 (v1 or v2)

300

322

No

MiniSeq

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

For any feedback or questions regarding this article (Illumina Knowledge Article #7400), contact Illumina Technical Support techsupport@illumina.com.

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