Multiple displacement amplification for malaria parasite DNA.

Multiple displacement amplification (MDA) using Phi29 has proved to be an efficient, high-fidelity method for whole genome amplification in many organisms. This project was designed to evaluate this approach for use with the malaria parasite Plasmodium falciparum. In particular, we were concerned th...

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Main Authors: Wang, Y, Nair, S, Nosten, F, Anderson, T
Format: Journal article
Language:English
Published: 2009
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author Wang, Y
Nair, S
Nosten, F
Anderson, T
author_facet Wang, Y
Nair, S
Nosten, F
Anderson, T
author_sort Wang, Y
collection OXFORD
description Multiple displacement amplification (MDA) using Phi29 has proved to be an efficient, high-fidelity method for whole genome amplification in many organisms. This project was designed to evaluate this approach for use with the malaria parasite Plasmodium falciparum. In particular, we were concerned that the AT richness and presence of contaminating human DNA could limit efficiency of MDA in this system. We amplified 60 DNA samples using phi29 and scored 14 microsatellites, 9 single-nucleotide polymorphisms (SNPs), and gene copy number at GTP-cyclohydrolase I both before and after MDA. We observed 100% concordance in 829 microsatellite genotypes and in 499 SNP genotypes. Furthermore, copy number estimates for the GTP-cyclohydrolase I gene were correlated (r(2) = 0.67) in pre- and postamplification samples. These data confirm that MDA permits scoring of a range of different types of polymorphisms in P. falciparum malaria and can be used to extend the life of valuable DNA stocks.
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spelling oxford-uuid:f6191aa3-1625-470c-87e0-0dff86a027912022-03-27T12:32:28ZMultiple displacement amplification for malaria parasite DNA.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:f6191aa3-1625-470c-87e0-0dff86a02791EnglishSymplectic Elements at Oxford2009Wang, YNair, SNosten, FAnderson, TMultiple displacement amplification (MDA) using Phi29 has proved to be an efficient, high-fidelity method for whole genome amplification in many organisms. This project was designed to evaluate this approach for use with the malaria parasite Plasmodium falciparum. In particular, we were concerned that the AT richness and presence of contaminating human DNA could limit efficiency of MDA in this system. We amplified 60 DNA samples using phi29 and scored 14 microsatellites, 9 single-nucleotide polymorphisms (SNPs), and gene copy number at GTP-cyclohydrolase I both before and after MDA. We observed 100% concordance in 829 microsatellite genotypes and in 499 SNP genotypes. Furthermore, copy number estimates for the GTP-cyclohydrolase I gene were correlated (r(2) = 0.67) in pre- and postamplification samples. These data confirm that MDA permits scoring of a range of different types of polymorphisms in P. falciparum malaria and can be used to extend the life of valuable DNA stocks.
spellingShingle Wang, Y
Nair, S
Nosten, F
Anderson, T
Multiple displacement amplification for malaria parasite DNA.
title Multiple displacement amplification for malaria parasite DNA.
title_full Multiple displacement amplification for malaria parasite DNA.
title_fullStr Multiple displacement amplification for malaria parasite DNA.
title_full_unstemmed Multiple displacement amplification for malaria parasite DNA.
title_short Multiple displacement amplification for malaria parasite DNA.
title_sort multiple displacement amplification for malaria parasite dna
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AT nairs multipledisplacementamplificationformalariaparasitedna
AT nostenf multipledisplacementamplificationformalariaparasitedna
AT andersont multipledisplacementamplificationformalariaparasitedna