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Biotechniques. 2015 May 1;58(5):253-7. doi: 10.2144/000114288. eCollection 2015 May.

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

Author information

1
Phyloinformatics Unit, RIKEN Center for Life Science Technologies, Kobe, Japan.

Abstract

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.

KEYWORDS:

CEGMA; genome assembly; genome scaffolding; mate pair library; tagmentation

PMID:
25967904
DOI:
10.2144/000114288
[Indexed for MEDLINE]
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