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DNA extraction techniques for DNA barcoding of minute gall‐inhabiting wasps
Author(s) -
DITTRICHSCHRÖDER GUDRUN,
WINGFIELD MICHAEL J.,
KLEIN HILDEGARD,
SLIPPERS BERNARD
Publication year - 2012
Publication title -
molecular ecology resources
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.96
H-Index - 136
eISSN - 1755-0998
pISSN - 1755-098X
DOI - 10.1111/j.1755-0998.2011.03074.x
Subject(s) - biology , gall , dna extraction , dna , botany , parasitoid , larva , agarose gel electrophoresis , dna barcoding , mitochondrial dna , extraction (chemistry) , zoology , polymerase chain reaction , chromatography , gene , genetics , chemistry
DNA extraction from minute hymenopterans and their larvae is difficult and challenging because of their small size indicating a low amount of starting material. Hence, 11 DNA extraction methods were compared to determine their efficacy in isolating DNA. Success of each method was scored on a 2% agarose gel after PCR of the cox 1 mitochondrial locus. A silica‐membrane‐based approach was the most successful, followed by a method using a combination of incubation buffers and a method using magnetic beads. The method using buffers was the most cost‐ and time effective. Using this method, larvae from Eucalyptus seed capsule galls could be assigned a role (parasitoid, gall former or inquiline) in the gall‐inhabiting complex.