Using long ssDNA polynucleotides to amplify STRs loci in degraded DNA samples
Publication Date
November 03, 2017
Authors
Martín E. Mautner, Agustín Pérez Santángelo, Rodrigo M. Corti Bielsa, Andrea Sala, et al
Volume
12
Issue
11
Pages
e0187190
DOI
https://dx.plos.org/10.1371/journal.pone.0187190
Publisher URL
http://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0187190
Scopus
85032894830
Mendeley
http://www.mendeley.com/research/using-long-ssdna-polynucleotides-amplify-strs-loci-degraded-dna-samples
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Mendeley | Further Information

{"title"=>"Using long ssDNA polynucleotides to amplify STRs loci in degraded DNA samples", "type"=>"journal", "authors"=>[{"first_name"=>"Martín E.", "last_name"=>"Mautner", "scopus_author_id"=>"57195953769"}, {"first_name"=>"Agustín Pérez", "last_name"=>"Santángelo", "scopus_author_id"=>"57196421599"}, {"first_name"=>"Rodrigo M.", "last_name"=>"Corti Bielsa", "scopus_author_id"=>"57196422254"}, {"first_name"=>"Andrea", "last_name"=>"Sala", "scopus_author_id"=>"7101706732"}, {"first_name"=>"Santiago", "last_name"=>"Ginart", "scopus_author_id"=>"56946809600"}, {"first_name"=>"Daniel", "last_name"=>"Corach", "scopus_author_id"=>"6701340533"}], "year"=>2017, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "pui"=>"619091742", "doi"=>"10.1371/journal.pone.0187190", "sgr"=>"85032894830", "isbn"=>"1111111111", "scopus"=>"2-s2.0-85032894830"}, "id"=>"0396f98c-44e1-3537-92e0-21a57dd2b23f", "abstract"=>"© 2017 Mautner et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Obtaining informative short tandem repeat (STR) profiles from degraded DNA samples is a challenging task usually undermined by locus or allele dropouts and peak-high imbalances observed in capillary electrophoresis (CE) electropherograms, especially for those markers with large amplicon sizes. We hereby show that the current STR assays may be greatly improved for the detection of genetic markers in degraded DNA samples by using long single stranded DNA polynucleotides (ssDNA polynucleotides) as surrogates for PCR primers. These long primers allow a closer annealing to the repeat sequences, thereby reducing the length of the template required for the amplification in fragmented DNA samples, while at the same time rendering amplicons of larger sizes suitable for multiplex assays. We also demonstrate that the annealing of long ssDNA polynucleotides does not need to be fully complementary in the 5’ region of the primers, thus allowing for the design of practically any long primer sequence for developing new multiplex assays. Furthermore, genotyping of intact DNA samples could also benefit from utilizing long primers since their close annealing to the target STR sequences may overcome wrong profiling generated by insertions/deletions present between the STR region and the annealing site of the primers. Additionally, long ssDNA polynucleotides might be utilized in multiplex PCR assays for other types of degraded or fragmented DNA, e.g. circulating, cell-free DNA (ccfDNA).", "link"=>"http://www.mendeley.com/research/using-long-ssdna-polynucleotides-amplify-strs-loci-degraded-dna-samples", "reader_count"=>6, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>3, "Researcher"=>1, "Other"=>2}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>3, "Researcher"=>1, "Other"=>2}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>4, "Agricultural and Biological Sciences"=>2}, "reader_count_by_subdiscipline"=>{"Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>2}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>4}}, "group_count"=>0}

Scopus | Further Information

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