Data from: Rapid identification of chloroplast haplotypes using High-Resolution Melting analysis
We have evaluated High Resolution Melt (HRM) analysis as a method for one-step haplotype identification in phylogeographic analysis. Using two adjoined internal amplicons (c.360 and 390bp) at the chloroplast rps16 intron (c.750bp) we applied HRM to identify haplotypes in 21 populations of two Europe...
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Online Access: | https://doi.org/10.5061/dryad.s904k |
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fttriple:oai:gotriple.eu:50|dedup_wf_001::1763289ce018fc8419608cd9e5dcbd60 2023-05-15T15:15:03+02:00 Data from: Rapid identification of chloroplast haplotypes using High-Resolution Melting analysis Dang, Xiao-Dong Howard-Williams, Emma Kelleher, Colin T. Meade, Conor V. 2012-01-01 https://doi.org/10.5061/dryad.s904k undefined unknown Dryad Digital Repository https://dx.doi.org/10.5061/dryad.s904k http://dx.doi.org/10.5061/dryad.s904k lic_creative-commons 10.5061/dryad.s904k oai:easy.dans.knaw.nl:easy-dataset:81403 oai:services.nod.dans.knaw.nl:Products/dans:oai:easy.dans.knaw.nl:easy-dataset:81403 10|openaire____::081b82f96300b6a6e3d282bad31cb6e2 re3data_____::r3d100000044 10|openaire____::9e3be59865b2c1c335d32dae2fe7b254 10|re3data_____::94816e6421eeb072e7742ce6a9decc5f 10|re3data_____::84e123776089ce3c7a33db98d9cd15a8 10|eurocrisdris::fe4903425d9040f680d8610d9079ea14 10|opendoar____::8b6dd7db9af49e67306feb59a8bdc52c Angiosperms Phylogeography Population Genetics - Empirical Speciation Conservation Genetics Europe Pleistocene Arenaria ciliata Arenaria norvegica Life sciences medicine and health care geo envir Dataset https://vocabularies.coar-repositories.org/resource_types/c_ddb1/ 2012 fttriple https://doi.org/10.5061/dryad.s904k 2023-01-22T17:23:37Z We have evaluated High Resolution Melt (HRM) analysis as a method for one-step haplotype identification in phylogeographic analysis. Using two adjoined internal amplicons (c.360 and 390bp) at the chloroplast rps16 intron (c.750bp) we applied HRM to identify haplotypes in 21 populations of two European arctic-alpine herb species Arenaria ciliata and A. norvegica (Caryophyllaceae). From 446 accessions studied, 20 composite rps16 haplotypes were identified by the melt-curve protocol, 18 of which could be identified uniquely. In a comparative sensitivity analysis with in silico PCR-RFLP, only 7 of these 20 haplotypes could be identified uniquely. Observed in vitro experimental HRM profiles were corroborated by in silico HRM analysis generated on uMeltSM. In silico mutation analysis carried out on a 360bp wild-type rps16I amplicon determined that the expected rate of missed SNP detection in vitro was similar to existing evaluations of HRM sensitivity, with transversion SNPs being more likely to go undetected compared to transition SNPs. Overall, pairwise melt peak differences between haplotypes were significantly correlated with genetic distance, and in vitro HRM successfully discriminated between all amplicon templates differing by 2 or more base changes (352 cases) and between 11 pairs of amplicons where the only difference was a single transition or transversion SNP. Only one pairwise comparison yielded no discernable HRM curve difference between haplotypes, these differed by one transversion (C/G) SNP. HRM analysis represents an untapped resource in phylogeographic analysis, and with appropriate primer design any polymorphic locus is potentially amenable to this single-reaction method for haplotype identification. Dang et al HRM Aligned rps16 sequencesAligned rps16 sequences for 20 haplotypes of Arenaria ciliata and A. norvegica (C01-c18, and N01-N02 resepctively). Alignment presented in fasta format and saved as .txt. Dataset Arctic Unknown Arctic |
institution |
Open Polar |
collection |
Unknown |
op_collection_id |
fttriple |
language |
unknown |
topic |
Angiosperms Phylogeography Population Genetics - Empirical Speciation Conservation Genetics Europe Pleistocene Arenaria ciliata Arenaria norvegica Life sciences medicine and health care geo envir |
spellingShingle |
Angiosperms Phylogeography Population Genetics - Empirical Speciation Conservation Genetics Europe Pleistocene Arenaria ciliata Arenaria norvegica Life sciences medicine and health care geo envir Dang, Xiao-Dong Howard-Williams, Emma Kelleher, Colin T. Meade, Conor V. Data from: Rapid identification of chloroplast haplotypes using High-Resolution Melting analysis |
topic_facet |
Angiosperms Phylogeography Population Genetics - Empirical Speciation Conservation Genetics Europe Pleistocene Arenaria ciliata Arenaria norvegica Life sciences medicine and health care geo envir |
description |
We have evaluated High Resolution Melt (HRM) analysis as a method for one-step haplotype identification in phylogeographic analysis. Using two adjoined internal amplicons (c.360 and 390bp) at the chloroplast rps16 intron (c.750bp) we applied HRM to identify haplotypes in 21 populations of two European arctic-alpine herb species Arenaria ciliata and A. norvegica (Caryophyllaceae). From 446 accessions studied, 20 composite rps16 haplotypes were identified by the melt-curve protocol, 18 of which could be identified uniquely. In a comparative sensitivity analysis with in silico PCR-RFLP, only 7 of these 20 haplotypes could be identified uniquely. Observed in vitro experimental HRM profiles were corroborated by in silico HRM analysis generated on uMeltSM. In silico mutation analysis carried out on a 360bp wild-type rps16I amplicon determined that the expected rate of missed SNP detection in vitro was similar to existing evaluations of HRM sensitivity, with transversion SNPs being more likely to go undetected compared to transition SNPs. Overall, pairwise melt peak differences between haplotypes were significantly correlated with genetic distance, and in vitro HRM successfully discriminated between all amplicon templates differing by 2 or more base changes (352 cases) and between 11 pairs of amplicons where the only difference was a single transition or transversion SNP. Only one pairwise comparison yielded no discernable HRM curve difference between haplotypes, these differed by one transversion (C/G) SNP. HRM analysis represents an untapped resource in phylogeographic analysis, and with appropriate primer design any polymorphic locus is potentially amenable to this single-reaction method for haplotype identification. Dang et al HRM Aligned rps16 sequencesAligned rps16 sequences for 20 haplotypes of Arenaria ciliata and A. norvegica (C01-c18, and N01-N02 resepctively). Alignment presented in fasta format and saved as .txt. |
format |
Dataset |
author |
Dang, Xiao-Dong Howard-Williams, Emma Kelleher, Colin T. Meade, Conor V. |
author_facet |
Dang, Xiao-Dong Howard-Williams, Emma Kelleher, Colin T. Meade, Conor V. |
author_sort |
Dang, Xiao-Dong |
title |
Data from: Rapid identification of chloroplast haplotypes using High-Resolution Melting analysis |
title_short |
Data from: Rapid identification of chloroplast haplotypes using High-Resolution Melting analysis |
title_full |
Data from: Rapid identification of chloroplast haplotypes using High-Resolution Melting analysis |
title_fullStr |
Data from: Rapid identification of chloroplast haplotypes using High-Resolution Melting analysis |
title_full_unstemmed |
Data from: Rapid identification of chloroplast haplotypes using High-Resolution Melting analysis |
title_sort |
data from: rapid identification of chloroplast haplotypes using high-resolution melting analysis |
publisher |
Dryad Digital Repository |
publishDate |
2012 |
url |
https://doi.org/10.5061/dryad.s904k |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
10.5061/dryad.s904k oai:easy.dans.knaw.nl:easy-dataset:81403 oai:services.nod.dans.knaw.nl:Products/dans:oai:easy.dans.knaw.nl:easy-dataset:81403 10|openaire____::081b82f96300b6a6e3d282bad31cb6e2 re3data_____::r3d100000044 10|openaire____::9e3be59865b2c1c335d32dae2fe7b254 10|re3data_____::94816e6421eeb072e7742ce6a9decc5f 10|re3data_____::84e123776089ce3c7a33db98d9cd15a8 10|eurocrisdris::fe4903425d9040f680d8610d9079ea14 10|opendoar____::8b6dd7db9af49e67306feb59a8bdc52c |
op_relation |
https://dx.doi.org/10.5061/dryad.s904k http://dx.doi.org/10.5061/dryad.s904k |
op_rights |
lic_creative-commons |
op_doi |
https://doi.org/10.5061/dryad.s904k |
_version_ |
1766345435603009536 |