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 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.
Wang
Y
Y
Southwest Foundation for Biomedical Research, San Antonio, Texas 78245, USA.
Nair
S
S
Nosten
F
F
Anderson
T J C
TJ
D000818:Animals; D016054:DNA, Protozoan / Q000097:blood / Q000737:chemistry*; D018900:Dinucleotide Repeats / Q000235:genetics; D006136:GTP Cyclohydrolase / Q000235:genetics; D005838:Genotype; D006801:Humans; D018895:Microsatellite Repeats / Q000235:genetics; D021141:Nucleic Acid Amplification Techniques / Q000379:methods* / Q000592:standards; D010963:Plasmodium falciparum / Q000145:classification / Q000201:enzymology / Q000235:genetics*; D011110:Polymorphism, Genetic / Q000235:genetics; D020641:Polymorphism, Single Nucleotide / Q000235:genetics
D016054:DNA, Protozoan; D006136:GTP Cyclohydrolase