BECAS
ANADON MarÍa Del Rosario
artículos
Título:
A Modified Combination of Two Methods for Isolation of Polymorphonuclear Granulocytes from Human Blood for Polymerase Chain Reaction (PCR)
Autor/es:
ALFREDO MARTÍNEZ; ROBERT PRIZONT; ROSARIO ANADÓN; FEDERICO KLOSOWSKI
Revista:
CLINICAL LABORATORY.
Editorial:
CLIN LAB PUBL
Referencias:
Año: 2017
ISSN:
1433-6510
Resumen:
Background: The aim of the study was to determine a rapid and simple method for isolation of polymorphonuclear(PMN) granulocytes from human blood appropriate for PCR of extracted DNA.Methods: A modification of the combined dextran plus Fycoll-Hypaque methods was the final method used in 41samples of normal white cells from humans. The changes encompassed a lower dextran-blood ratio (2:1), a shortercentrifugation time of leukocytes, a small volume of Ficoll-Hypaque suspension, and a decreased speed in thelast centrifugation. The final key point was skipping the lysis of erythrocytes to avoid interference by salt solutions.Results: These modifications rendered a mean PMN granulocyte recovery and purity of over 95% with a very narrow2 - 8% coefficient of variation and did not alter the purity of DNA extraction.Conclusions: The combined dextran plus Fycoll-Hypaque method changes offer a fast and simple method to obtainan adequate purity of granulocytes for PCR.
