title: A simple, fast, and inexpensive CTAB-PVP-silica based method for genomic DNA isolation from single, small insect larvae and pupae.
abstract: In this study, we report a modified CTAB-PVP method combined with silicon dioxide (silica) treatment for the extraction of high quality genomic DNA from a single larva or pupa. This method efficiently obtains DNA from small specimens, which is difficult and challenging because of the small amount of starting tissue. Maceration with liquid nitrogen, phenol treatment, and the ethanol precipitation step are eliminated using this methodology. The A260/A280 absorbance ratios of the isolated DNA were approximately 1.8, suggesting that the DNA is pure and can be used for further molecular analysis. The quality of the isolated DNA permits molecular applications and represents a fast, cheap, and effective alternative method for laboratories with low budgets.
authors.affiliation: Laboratorio de Biología Molecular de Plantas, Departamento de Producción Agrícola, Facultad de Ciencias Agronómicas, Universidad de Tarapacá, Arica, Chile whuanca@uta.cl.
authors.forename: W
authors.initials: W
authors.lastname: Huanca-Mamani
authors.affiliation: Laboratorio de Biología Molecular de Plantas, Departamento de Producción Agrícola, Facultad de Ciencias Agronómicas, Universidad de Tarapacá, Arica, Chile.
authors.forename: D
authors.initials: D
authors.lastname: Rivera-Cabello
authors.affiliation: Laboratorio de Biología Molecular de Plantas, Departamento de Producción Agrícola, Facultad de Ciencias Agronómicas, Universidad de Tarapacá, Arica, Chile.
authors.forename: J
authors.initials: J
authors.lastname: Maita-Maita
mesh_terms: D000818:Animals; D001483:Base Sequence; D000077286:Cetrimonium; D002593:Cetrimonium Compounds / Q000737:chemistry*; D003362:Cost-Benefit Analysis; D004247:DNA / Q000302:isolation & purification*; D003576:Electron Transport Complex IV / Q000235:genetics; D049750:Genome, Insect; D007313:Insecta / Q000235:genetics*; D007814:Larva / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000191:economics* / Q000379:methods*; D011205:Povidone / Q000737:chemistry*; D011679:Pupa / Q000235:genetics*; D017422:Sequence Analysis, DNA; D012822:Silicon Dioxide / Q000737:chemistry*
chemical_list: D002593:Cetrimonium Compounds; D012822:Silicon Dioxide; D004247:DNA; D003576:Electron Transport Complex IV; D011205:Povidone; D000077286:Cetrimonium