Optimization and cost-saving in tagmentation-based mate-pair library preparation and sequencing

BioTechniques

Kazu Itomi¹  Shigehiro Kuraku¹  Kaori Tatsumi¹  Chiharu Tanegashima¹  Osamu Nishimura¹ 

[1] 1Phyloinformatics Unit, RIKEN Center for Life Science Technologies, Kobe, Japan;

关键词: mate pair library;    genome assembly;    genome scaffolding;    CEGMA;    tagmentation;   

DOI  :  10.2144/000114288

来源: DOAJ

【 摘 要 】

In de novo genome sequencing, mate-pair reads are crucial for scaffolding assembled contigs. However, preparation of mate-pair libraries is not a trivial task, even when using one of the latest approaches, the Nextera Mate Pair Sample Prep Kit from Illumina. To reduce cost and enhance library yield and fidelity when using this kit, we have modified the manufacturer's protocol based on (i) variable tagmentation conditions, (ii) intensive DNA shearing to decrease library insert length, and (iii) sequencing on an Illumina HiSeq with >150 cycles. Finally, we provide additional suggestions for further improvement in the application of this kit.

【 授权许可】

Unknown