TY - JOUR
T1 - Brugia malayi
T2 - Whole genome amplification for genomic characterization of filarial parasites
AU - McNulty, Samantha N.
AU - Weil, Gary J.
AU - Heinz, Michael
AU - Crosby, Seth D.
AU - Fischer, Peter U.
PY - 2008/6
Y1 - 2008/6
N2 - Genetic characterization of field isolates and clinical specimens of filarial nematodes is often limited by a shortage of DNA; therefore, we evaluated a multiple displacement amplification (MDA) based whole genome amplification method. The quality of amplified DNA was examined by conventional PCR, real-time PCR, and DNA hybridization. MDA of 5.0 ng of adult Brugia malayi DNA and one-fifteenth of the DNA isolated from a single microfilaria resulted in 6.3 and 4.2 μg of amplified DNA, respectively. Amplified DNA was equivalent to native genomic DNA for hybridization to B. malayi BAC library clones or to an oligonucleotide microarray with approximately 18,000 filarial DNA sequences. MDA is useful for whole genome amplification of filarial DNA from very small amounts of starting material. This technology will permit detailed studies of genetic diversity that were not previously feasible.
AB - Genetic characterization of field isolates and clinical specimens of filarial nematodes is often limited by a shortage of DNA; therefore, we evaluated a multiple displacement amplification (MDA) based whole genome amplification method. The quality of amplified DNA was examined by conventional PCR, real-time PCR, and DNA hybridization. MDA of 5.0 ng of adult Brugia malayi DNA and one-fifteenth of the DNA isolated from a single microfilaria resulted in 6.3 and 4.2 μg of amplified DNA, respectively. Amplified DNA was equivalent to native genomic DNA for hybridization to B. malayi BAC library clones or to an oligonucleotide microarray with approximately 18,000 filarial DNA sequences. MDA is useful for whole genome amplification of filarial DNA from very small amounts of starting material. This technology will permit detailed studies of genetic diversity that were not previously feasible.
KW - Brugia malayi
KW - Cycle threshold (C)
KW - Microarray
KW - Multiple displacement amplification (MDA)
KW - Oligonucleotide microarray
KW - Real-time PCR
KW - Whole genome amplification (WGA)
UR - http://www.scopus.com/inward/record.url?scp=42749087622&partnerID=8YFLogxK
U2 - 10.1016/j.exppara.2008.02.003
DO - 10.1016/j.exppara.2008.02.003
M3 - Article
C2 - 18359019
AN - SCOPUS:42749087622
SN - 0014-4894
VL - 119
SP - 256
EP - 263
JO - Experimental Parasitology
JF - Experimental Parasitology
IS - 2
ER -