DNA extraction from archival Giemsa-stained bone-marrow slides: comparison of six rapid methods.
The ability of six rapid DNA extraction procedures to provide DNA for the polymerase chain reaction from archival Giemsa-stained bone marrow slides was tested on 120 samples. Boiling in distilled water, freeze-thaw method, boiling in 10% Chelex-100 resin solution, proteinase K/Tween 20/NP-40 method coupled with simplified phenol/ chloroform/isoamyl alcohol protocol or salting-out procedure using saturated NaCl and modification of commercial QIAamp procedure (Qiagen. Chatsworth, Calif.) gave DNA extraction efficiencies of 50%, 70%, 85%, 95%, 100% and 100%, respectively. Our results demonstrate that rough DNA extraction methods have decreased efficiencies compared to complete DNA extraction protocols and that the latter are required to ensure highly reproducible results from archival Giemsa-stained bone marrow slides.
Vince
A
A
Dr Fran Mihaljević University Hospital for Infectious Diseases, Zagreb, Croatia.
Poljak
M
M
Seme
K
K
D001399:Azure Stains; D001854:Bone Marrow Cells / Q000737:chemistry; D004247:DNA / Q000302:isolation & purification*; D016591:Histocytological Preparation Techniques; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods
D001399:Azure Stains; D004247:DNA