Fecal sample collection and DNA extraction
We collected fecal samples from 41 individuals (3-15 samples per
individual) living in 14 family groups, including 19 putative offspring
of 9 family groups (1 to 5 offspring per group). Five other groups
either did not have offspring during the study period (or they had
disappeared before we could collect samples) or the samples could not be
collected because the offspring were still very small and thus their
defecations too diminutive to be detected or not habituated enough. Also
due to differential habituation to the presence of humans, for some
groups we could not obtain samples from all group members. For those
groups that were habituated in the beginning of the study period, we
collected samples from offspring from several consecutive years.
Fecal samples were collected immediately after an identified individual
was seen defecating. We kept the samples in 15 mL falcon tubes
containing silica gel beads (Carl Roth, Karlsruhe, Germany) and stored
them at ambient temperature, replacing the silica beads when necessary,
until shipping to the Germany.
We extracted DNA (at least two samples per individual) from ca. 200 mg
of feces using the Macherey-Nagel NucleSpin DNA stool kit with a final
elution of the DNA in 50 µL elution buffer. DNA concentration of the
extracts was measured using NanoDrop (Spectrophotometer ND-1000, PEQLAB
Biotechnologie GmbH) and Qubit Fluorometer (Thermo Fisher).