Optimized construction of microsatellite-enriched libraries.

Abstract:

:The construction of microsatellite-enriched libraries is an indispensable tool to search for molecular markers as complete genome sequences are still not available for the majority of species of interest. Numerous protocols are available in the literature for the construction of these libraries; however, sometimes their low efficiency or lack of optimization in the protocols can restrict their efficacy. We have designed and tested various adapters and ligation methods; we also tested oligo-repeat combinations and hybridization temperatures, and created libraries with this new protocol for four organisms: Ipomoea batatas (L.) Lam, Chionanthus retusus Lindley & Paxton, Rotylenchulus reniformis Linford & Olivera and Puccinia kuehnii W. Krüger. The number of microsatellites detected for these species ranged from 2494 to 3919 per Mb of nonredundant sequence, that was 0.86 and 1.53 microsatellites per contig, with 37-66% of di-nucleotide motifs and 21-49% of tri- to octa-nucleotide repeats combined. A simplified protocol is provided for the successful generation of SSR-enriched libraries.

journal_name

Mol Ecol Resour

authors

Techen N,Arias RS,Glynn NC,Pan Z,Khan IA,Scheffler BE

doi

10.1111/j.1755-0998.2009.02802.x

subject

Has Abstract

pub_date

2010-05-01 00:00:00

pages

508-15

issue

3

eissn

1755-098X

issn

1755-0998

journal_volume

10

pub_type

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