Isolation of high-molecular-weight DNA from small samples of blood having nucleated erythrocytes, collected, transported, and stored at room temperature.
Blood samples collected in the field for isolating DNA suitable for molecular analysis need special care in their storage and handling. In this article, we describe a simple method for the isolation of good-quality high-molecular-weight DNA that does not require low temperature conditions during collection, storage, and/or transportation of blood samples. This method involves smearing small aliquots of blood onto clean slides and air drying them at room temperature. The slides with blood smears can then be transported or stored at room temperature and still serve as a very good source of high-molecular-weight DNA. Genomic DNA from these samples can be extracted by organic phase separation (phenol-chloroform extraction) after lysis. The DNA thus obtained is of high quality and yields DNA fingerprints qualitatively similar to those prepared from corresponding control DNA isolated from frozen blood samples. Needing minimal facilities at field sites, the method is very convenient for conducting RFLP analysis of wild/field populations for demographic, behavioral, and ecologic studies.
Centre for Cellular and Molecular Biology, Hyderabad, India.
R K
RK
Aggarwal
J W
JW
Lang
L
L
Singh
D000489:Alligators and Crocodiles / Q000097:blood; D000818:Animals; D001800:Blood Specimen Collection; D004247:DNA / Q000097:blood* / Q000302:isolation & purification; D016172:DNA Fingerprinting; D005399:Fishes / Q000097:blood; D012150:Polymorphism, Restriction Fragment Length; D013696:Temperature
D004247:DNA