Isolation of high-molecular-weight DNA from insects.
A simple and rapid method for the isolation of high-molecular-weight DNA from insects is described. The method does not require CsCl ultracentrifugation or extensive dialysis. High-molecular-weight DNA was obtained within 24 h. Since the entire insect was used for DNA isolation, an initial nuclei-enriched fraction was required. Genomic DNA was extracted from lysed nuclei by organic phase separation (liquid/liquid extraction). This method has been successfully applied to the isolation and purification of DNA from eight different adult insects (Heliothis zea, Musca autumnalis, M. domestica, Blatta orientalis, Tenebrio molitor, Lymantria dispar, Ostrinia nubilalis, and Manduca sexta). The recovered DNA can be cleaved with restriction endonucleases, ligated efficiently using standard cloning vectors, and hybridized to synthetic oligonucleotides.
Agricultural Research Service, USDA, Beltsville, Maryland 20705.
J M
JM
Henry
A K
AK
Raina
R L
RL
Ridgway
D000818:Animals; D015139:Blotting, Southern; D002458:Cell Fractionation; D002467:Cell Nucleus / Q000032:analysis; D004247:DNA / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D007313:Insecta / Q000032:analysis* / Q000235:genetics; D008722:Methods; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization; D012313:RNA / Q000032:analysis
D012313:RNA; D004247:DNA