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Making the first step: practical considerations for the isolation of low‐copy nuclear sequence markers
Author(s) -
Schlüter Philipp M.,
Stuessy Tod F.,
Paulus Hannes F.
Publication year - 2005
Publication title -
taxon
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.819
H-Index - 81
eISSN - 1996-8175
pISSN - 0040-0262
DOI - 10.2307/25065432
Subject(s) - biology , sequence (biology) , genetics , computational biology , dna sequencing , isolation (microbiology) , nuclear dna , cloning (programming) , nuclear gene , population , dna , organism , sequence analysis , complementary dna , gene , genome , bioinformatics , mitochondrial dna , computer science , demography , sociology , programming language
Abstract In many plant groups, the use of low‐copy nuclear sequence markers for phylogenetics and population genetics has been hindered by their limited availability. Although it may be possible to PCR amplify low‐copy markers using primers designed for use with other plant groups, this does not always yield the desired results. Here, we suggest several alternative approaches to begin the isolation and characterisation of novel low‐copy markers when there is little or no sequence information available. These alternatives are: (1) the design of new primers from information in the sequence databases; (2) isolation of homologous DNA using a gene probe from another organism; (3) characterisation of sequence markers from DNA fingerprints; and (4) obtaining novel sequences via cDNA cloning.