PCR of immobilized DNA molecules isolated from human blood cells by a non-enzymatic method.
DNA molecules suitable for amplification by Polymerase Chain Reaction were obtained by immobilizing whole blood or isolated leukocytes and incubating the immobilized cells for one hour with the known non-enzymatic solution described for preparing intact DNA molecules for PFGE. Cell immobilization was done in agarose gels and punches of 1.2 mm of diameter had the amount of DNA needed for amplifying chromosomal and mitochondrial sequences, many times. The approach was successfully used in preparing DNA molecules from multiple samples in flat-bottom 96-well ELISA plates. The procedure is simple and does not demand special conditions for sample transportation or conservation; thus, it should be useful to collect and process samples under field conditions in epidemiological studies.
Suárez
Viana Manrique
VM
Molecular Biology Department, Cuban Neurosciences Center, Havana, Cuba.
Clarke
David Higginson
DH
López-Canovas
Lilia
L
Riverón
Ana María
AM
D001483:Base Sequence; D015374:Biosensing Techniques; D001773:Blood Cells / Q000737:chemistry / Q000378:metabolism*; D002877:Chromosomes, Human / Q000235:genetics; D004247:DNA / Q000097:blood* / Q000235:genetics / Q000302:isolation & purification*; D004272:DNA, Mitochondrial / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D006801:Humans; D021141:Nucleic Acid Amplification Techniques; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D004272:DNA, Mitochondrial; D004247:DNA