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A factorial design experiment as a pilot study for noninvasive genetic sampling
Author(s) -
RENAN SHARON,
SPEYER EDITH,
SHAHAR NAAMA,
GUETA TOMER,
TEMPLETON ALAN R.,
BARDAVID SHIRLI
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.1755-0998.2012.03170.x
Subject(s) - biology , dna extraction , microsatellite , sampling (signal processing) , factorial experiment , computational biology , protocol (science) , microbiology and biotechnology , polymerase chain reaction , genetics , computer science , machine learning , pathology , medicine , allele , alternative medicine , filter (signal processing) , gene , computer vision
Noninvasive genetic sampling has increasingly been used in ecological and conservation studies during the last decade. A major part of the noninvasive genetic literature is dedicated to the search for optimal protocols, by comparing different methods of collection, preservation and extraction of DNA from noninvasive materials. However, the lack of quantitative comparisons among these studies and the possibility that different methods are optimal for different systems make it difficult to decide which protocol to use. Moreover, most studies that have compared different methods focused on a single factor – collection, preservation or extraction – while there could be interactions between these factors. We designed a factorial experiment, as a pilot study, aimed at exploring the effect of several collection, preservation and extraction methods, and the interactions between them, on the quality and amplification success of DNA obtained from Asiatic wild ass ( Equus hemionus ) faeces in Israel. The amplification success rates of one mitochondrial DNA and four microsatellite markers differed substantially as a function of collection, preservation and extraction methods and their interactions. The most efficient combination for our system integrated the use of swabs as a collection method with preservation at −20 °C and with the Qiagen DNA Stool Kit with modifications as the DNA extraction method. The significant interaction found between the collection, preservation methods and the extraction methods reinforces the importance of conducting a factorial design experiment, rather than examining each factor separately, as a pilot study before initiating a full‐scale noninvasive research project.