Amplification of human mitochondrial DNA (mtDNA) has been widely used in
population genetics, human evolutionary and molecular anthropology studies.
mtDNA hypervariable segments I and II (HVSI and HVSII) were shown to be a
suitable tool in genetic analyses due to the unique properties of mtDNA, such
as the lack of recombination, maternal mode of inheritance, rapid
evolutionary rate and high population-specific polymorphisms. Here we present
a rapid and low-cost method for direct PCR amplification of a 330 bp fragment
of HVSI from buccal cell samples. Avoiding the DNA isolation step makes this
method appropriate for the analysis of a large number of samples in a short
period of time. Since the transportation of samples and fieldwork conditions
can affect the quality of samples and subsequent DNA analysis, we tested the
effects of long-term storage of buccal cell swabs on the suitability of such
samples for direct PCR amplification. We efficiently amplified a 330 bp
fragment of HVSI even after the long-term storage of buccal cells at room
temperature, +4?C or at -20?C, for up to eight months. All examined PCR
products were successfully sequenced, regardless of sample storage time and
conditions. Our results suggest that the direct PCR amplification of the HVSI
region from buccal cells is a method well suited for large-scale mtDNA
population studies.
Long-term Storage and Recovery of Buccal Cell DNA from Treated Cards
