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Efficiency of microsatellite enrichment inProsopis chilensis using magnetic capture

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Abstract

Microsatellites (i.e., simple sequence repeats [SSRs]) are highly variable genetic markers that are widely used at an intraspecific level in population genetic studies. Here we employed an enrichment strategy for microsatellite isolation by using microsatellite oligoprobes and magnetic capture of the fragments (Fischer and Bachmann, 1998) inProsopis chilensis (Mol.) Stuntz (Fabaceae). We analyzed the obtained level of enrichment by sequencing 120 enriched genomic fragments. A total of 521 SSR motives were detected. According to specific search criteria (SSR motifs ≥3 repeat units and ≥6 bp length), 95.8% of the clones contained SSR motifs. Of these, 7.8% showed homology to chloroplast sequences and 92.2% to nuclear sequences. When regarding only nuclear SSRs with 5 or more repeat units and a minimum length of 10 bp, the level of enrichment was 30.8%. A FASTA search against the European Molecular Biology Laboratory (EMBL) database univocally revealed 4 clones in transcribed regions, 102 clones in genomic regions with unknown function, and 9 clones in chloroplast regions. Among the loci with longer repeat units (≥10 bp, ≥5 repeat units), 3 were in transcribed regions and 65 were in other genomic regions. We discuss the applicability of these markers for population genetic studies.

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Abbreviations

SSR:

simple sequence repeat

References

  • Cordeiro GM, Maguire TL, Edwards KJ, and Henry RJ (1999) Optimization of a microsatellite enrichment technique inSaccharum spp. Plant Mol Biol Rep 17: 225–229.

    Article  CAS  Google Scholar 

  • Edwards KJ, Barker JHA, Daly A, Jones C, and Karp A (1996) Microsatellite libraries enriched for several microsatellite sequences in plants. BioTechniques 20: 758–760.

    PubMed  CAS  Google Scholar 

  • Fischer D and Bachman K (1998) Microsatellite enrichment in organisms with large genomes (Allium cepa L.). BioTechniques 24: 796–802.

    PubMed  CAS  Google Scholar 

  • Hamilton MB, Pincus EL, Di Fiore A, and Fleischer RC (1999) Universal linker and ligation procedures for construction of genomic DNA libraries enriched for microsatellites. BioTechniques 27: 500–507.

    PubMed  CAS  Google Scholar 

  • Jakse J and Javornik B (2001) High throughput isolation of microsatellites in hop (Humulus lupulus L.). Plant Mol Biol Rep 19: 217–226.

    Article  CAS  Google Scholar 

  • Jurka J and Pethiyagoda C (1995) Simple repetitive DNA sequences from primates: compilation and analysis. J Mol Evol 40: 120–126.

    Article  PubMed  CAS  Google Scholar 

  • Karagyozov L, Kalcheva ID, and Chapman VM (1993) Construction of random small-insert genomic libraries highly enriched for simple sequence repeats. Nucleic Acids Res 21: 3911–3912.

    Article  PubMed  CAS  Google Scholar 

  • Kijas JMH, Fowler JCS, Garbett CA, and Thomas MR (1994) Enrichment of microsatellites from the citrus genome using biotinylated oligonucleotide sequences bound to streptavidin-coated magnetic particles. BioTechniques 16: 657–662.

    Google Scholar 

  • Litt M and Luty JA (1989) A hypervariable microsatellite revealed by in vitro amplification of a dinucleotide repeat within the cardiac muscle actin gene. Am J Hum Genet 44: 397–401.

    PubMed  CAS  Google Scholar 

  • Marchuk D, Drumm M, Saulino A, and Collins FS (1990) Construction of T-vectors, a rapid and general system for direct cloning of unmodified PCR products. Nucleic Acids Res 19: 1154.

    Article  Google Scholar 

  • Morgante M, Hanafey M, and Powell W (2002) Microsatellites are preferentially associated with nonrepetitive DNA in plant genomes. Nat Genet 30: 194–200.

    Article  PubMed  CAS  Google Scholar 

  • Paetkau D (1999) Microsatellites obtained using strand extension: an enrichment protocol. BioTechniques 26: 690–697.

    PubMed  CAS  Google Scholar 

  • Pandey M, Gailing O, Fischer D, Hattemer HH, and Finkeldey R (2004) Characterization of microsatellite markers in sycamore (Acer pseudoplatanus L.). Mol Ecol Notes 4: 253–255.

    Article  CAS  Google Scholar 

  • Pearson WR and Lipman DJ (1988) Improved tools for biological sequence comparison. Proc Acad Nat Sci USA 85: 2444–2448.

    Article  CAS  Google Scholar 

  • Pearson WR (1990) Rapid and sensitive sequence comparison with FASTP and FASTA. Meth Enzymol 183: 63–98.

    Article  PubMed  CAS  Google Scholar 

  • Rodrigues NB, LoVerde PT, Romanha AJ, and Oliveira G (2002) Characterization of newSchistosoma mansoni microsatellite loci in sequences obtained from public DNA database and microsatellite enriched genomic libraries,Mem Inst Oswaldo Cruz, Rio de Janerio, vol 97 (Suppl I): 71–75.

    Article  CAS  Google Scholar 

  • Saneyoshi U, Yoshihiko T, and Izumi W (2003) Development of microsatellite markers inPrimula sieboldii E. Morren, a threatened Japanese perennial herb. Conserv Genet 4: 809–811.

    Article  Google Scholar 

  • Tautz D (1989) Hypervariability of simple sequences as a general source for polymorphic DNA markers. Nucleic Acids Res 17: 6463–6471.

    Article  PubMed  CAS  Google Scholar 

  • Watterson GA (1978) The homozygosity test of neutrality. Genetics 88: 405–417.

    PubMed  CAS  Google Scholar 

  • Weber JL and May PE (1989) Abundant class of human DNA polymorphism which can be typed using the polymerase chain reaction. Am J Hum Genet 44: 388–396.

    PubMed  CAS  Google Scholar 

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Correspondence to Oliver Gailing.

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Mottura, M.C., Gailing, O., Verga, A.R. et al. Efficiency of microsatellite enrichment inProsopis chilensis using magnetic capture. Plant Mol Biol Rep 22, 251–258 (2004). https://doi.org/10.1007/BF02773135

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