DNA extraction from fixed cytogenetic cell suspensions.
We developed a procedure for DNA extraction from small volumes of fixed cell suspensions previously prepared for conventional cytogenetic analysis. Good quality DNA was isolated with a fast and simple protocol using DNAzol reagent. This provided suitable DNA for various types of molecular analyses, including polymerase chain reaction, restriction fragment length polymorphism, denaturing high-performance liquid chromatography, and direct sequencing. This technique provides sufficient material for such test, which are important for diagnosis of neoplastic diseases in pediatric patients.
Amorim
M R
MR
Divisão de Medicina Experimental, Instituto Nacional de Câncer, Rio de Janeiro, RJ, Brasil.
Vargas
F R
FR
Llerena
J C
JC
Pombo-de-Oliveira
M S
MS
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003582:Cytogenetics / Q000379:methods*; D004247:DNA / Q000032:analysis*; D005091:Exons; D016678:Genome; D006801:Humans; D007621:Karyotyping; D009369:Neoplasms / Q000175:diagnosis / Q000235:genetics; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D017422:Sequence Analysis, DNA / Q000379:methods
D004247:DNA