文库定量与质量控制简明指引

测序文库精确的定量与准确的质量检测对于二代测序成功是十分关键的。对于不同测序文库试剂盒，我们推荐使用不同的定量和质量控制的方法。在本篇技术文档中，我们总结了目前所有Illlumina建库试剂盒的文库定量与质量控制的方法。在测序前，利用下表中的方法对最终文库进行鉴定。

DNA试剂盒-仅用于科研

试剂盒	定量	质量控制
TruSeq Nano DNA	荧光方法或qPCR	BioAnalyzer 或 Fragment Analyzer
TruSeq PCR Free	qPCR	BioAnalyzer 或 Fragment Analyzer
Illumina DNA Prep, (M) Tagmentation (formerly Nextera DNA Flex)	荧光方法(Qubit/PicoGreen)	BioAnalyzer 或 Fragment Analyzer
Nextera XT	荧光方法或基于磁珠均一化	BioAnalyzer (当使用荧光方法时需要；基于磁珠进行均一化时为可选)
Nextera Mate Pair	qPCR/BioAnalyzer	BioAnalyzer或琼脂糖凝胶
Illumina DNA Prep with Enrichment, (S) Tagmentation (formerly Nextera DNA Flex for Enrichment)	Qubit	BioAnalyzer
Illumina DNA PCR-Free Prep, Tagmentation	Qubit ssDNA或基于投入量的qPCR, 具体细节可参照 Reference Guide	N/A (最终文库为单链DNA)
TruSeq Exome	荧光方法或qPCR	BioAnalyzer 或 Fragment Analyzer
Nextera DNA Exome	荧光方法或qPCR	BioAnalyzer 或 Fragment Analyzer
Nextera Rapid Capture Custom Enrichment	荧光方法或qPCR	BioAnalyzer
TruSight Inherited Disease	荧光方法或qPCR	BioAnalyzer
TruSight Tumor 15	荧光方法(AccuClear/PicoGreen/Qubit)	BioAnalyzer或琼脂糖凝胶
TruSight Tumor 26 (TruSight Tumor)	BioAnalyzer	琼脂糖凝胶/BioAnalyzer
Ampliseq (DNA Panels)	BioAnalyzer/Fragment Analyzer/qPCR/Qubit/AccuClear/QuantIT	BioAnalyzer 或 Fragment Analyzer
TruSight Myeloid	基于磁珠均一化	BioAnalyzer或琼脂糖凝胶
TruSeq Amplicon Cancer Panel	基于磁珠均一化	BioAnalyzer或琼脂糖凝胶
TruSeq Bovine Parentage Sequencing Panel	基于磁珠均一化	BioAnalyzer/Fragment Analyzer/Agarose Gel
TruSeq ChIP	qPCR	BioAnalyzer
TruSeq Methyl Capture EPIC Library Prep Kit	Qubit	BioAnalyzer

RNA 试剂盒-仅用于科研

试剂盒	定量	质量控制
TruSeq RNA v2	qPCR	BioAnalyzer 或 Fragment Analyzer
TruSeq Stranded mRNA	qPCR	BioAnalyzer 或 Fragment Analyzer
TruSeq Stranded Total RNA	qPCR	BioAnalyzer 或 Fragment Analyzer
TruSeq RNA Exome	qPCR/荧光方法	BioAnalyzer 或 Fragment Analyzer
Illumina Stranded mRNA Prep, Ligation	Qubit	BioAnalyzer
Illumina Stranded Total RNA Prep with Ribo-Zero Plus	Qubit	BioAnalyzer
Illumina RNA Prep with Enrichment, (L) Tagmentation	Qubit	BioAnalyzer
TruSight RNA Pan-Cancer	qPCR	BioAnalyzer 或 Fragment Analyzer
TruSight RNA Fusion	qPCR	BioAnalyzer 或 Fragment Analyzer
TruSeq Targeted RNA Expression	BioAnalyzer 或 Fragment Analyzer	BioAnalyzer 或 Fragment Analyzer
TruSeq Small RNA	BioAnalyzer	BioAnalyzer
Ampiseq (RNA Panels)	BioAnalyzer/Fragment Analyzer/qPCR/Qubit/AccuClear/QuantIT	BioAnalyzer 或 Fragment Analyzer
SureCell WTA 3'	BioAnalyzer	BioAnalyzer

DNA和RNA试剂盒-仅用于科研

试剂盒	定量	质量控制
TruSight Tumor 170	Fluorometric Method/基于磁珠均一化	不需要
TruSight Oncology 500	AccuClear/基于磁珠均一化	不需要
TruSight Oncology 500 High Throughput	AccuClear/基于磁珠均一化	不需要
TruSight Oncology 500 ctDNA	AccuClear/基于磁珠均一化	不需要

分子诊断试剂盒

Kits	定量	质量控制
Cystic Fibrosis 139-Variant	基于磁珠均一化	不需要
Cystic Fibrosis Clinical Sequencing	基于磁珠均一化	不需要
TruSeq Custom Amplicon Dx	基于磁珠均一化	不需要

注意：

· 荧光方法为双链DNA特异荧光染料方法，例如QuBit, PicoGreen和AccuClear。

· qPCR定量方法更多的信息可以参考qPCR guide。

· 基于磁珠进行均一化的文库不需要进行质量控制。

For any feedback or questions regarding this article (Illumina Knowledge Article #7296), contact Illumina Technical Support [e-mail].