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 Resourjournal_title
Molecular ecology resourcesauthors
Techen N,Arias RS,Glynn NC,Pan Z,Khan IA,Scheffler BEdoi
10.1111/j.1755-0998.2009.02802.xsubject
Has Abstractpub_date
2010-05-01 00:00:00pages
508-15issue
3eissn
1755-098Xissn
1755-0998journal_volume
10pub_type
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