Empirical evaluation of preservation methods for faecal DNA.
We evaluate the relative effectiveness of four methods for preserving faecal samples for DNA analysis. PCR assays of fresh faecal samples collected from free-ranging baboons showed that amplification success was dependent on preservation method, PCR-product size, and whether nuclear or mitochondrial DNA was assayed. Storage in a DMSO/EDTA/Tris/salt solution (DETs) was most effective for preserving nuclear DNA, but storage in 70% ethanol, freezing at -20 degrees C and drying performed approximately equally well for mitochondrial DNA and short (< 200 bp) nuclear DNA fragments. Because faecal DNA is diluted and degraded, repeated extractions from faeces may be necessary and short nuclear markers should be employed for genotyping. A review of molecular scatology studies further suggests that three to six faeces per individual should be collected.
Frantzen
M A
MA
Department of Zoology and Entomology, University of Pretoria, South Africa. michaelk@biology.ucla.edu
Silk
J B
JB
Ferguson
J W
JW
Wayne
R K
RK
Kohn
M H
MH
D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics; D004272:DNA, Mitochondrial / Q000235:genetics / Q000302:isolation & purification; D017753:Ecosystem; D003576:Electron Transport Complex IV / Q000235:genetics; D005069:Evaluation Studies as Topic; D005243:Feces / Q000737:chemistry*; D005828:Genetics, Population; D005838:Genotype; D010215:Papio / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011309:Preservation, Biological / Q000379:methods*; D012996:Solutions
D017931:DNA Primers; D004272:DNA, Mitochondrial; D012996:Solutions; D004247:DNA; D003576:Electron Transport Complex IV