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Electrophoretic identification of new genomic profiles with a modified selective amplification of microsatellite polymorphic loci technique based on AT/AAT polymorphic repeats
Author(s) -
Palacios Gustavo,
Bustamante Silvia,
Molina Carlos,
Winter Peter,
Kahl Günter
Publication year - 2002
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(200210)23:19<3341::aid-elps3341>3.0.co;2-c
Subject(s) - microsatellite , biology , genome , primer (cosmetics) , genetics , computational biology , gene , physics , allele , thermodynamics
The present paper introduces improvements of the conventional selective amplification of microsatellite polymorphic loci (SAMPL) technique, that exploit AT‐rich microsatellite primers. Generally, AT/AAT microsatellites are frequent components of eukaryotic genomes, but their ubiquity and polymorphic information content (PIC) could not be exploited yet, because standard SAMPL conditions did not allow amplifications. Here we report (i) on the design of new versatile AT‐rich microsatellite primers, that are combined with (ii) a modified SAMPL adapter primer (called Eco RI‐Short), and (iii) special polymerase chain reaction (PCR) amplification regimes. The novel SAMPL procedure expands the range of useful microsatellite primers to AT‐rich sequences and produces a high number of bands and a clear banding pattern, and detects polymorphisms in otherwise nonpolymorphic genomes of plants ( Dioscorea alata, D. rotundata ) and a fungus ( Mycosphaerella fijiensis ).