Voltage-induced release of nucleic acids from palaeontological samples.
Most of the protocols for the recovery of ancient DNA from palaeontological specimens are time-consuming and tend to yield inconsistent polymerase chain reaction (PCR) results. "Voltage-induced release" is a novel and rapid approach for the extraction of ancient DNA. Nucleic acids are directly electrophoresed out of powder derived from hard and soft tissues. This technique is much faster than other methods in which pulverized tissue conventionally undergoes time-consuming crude lysis steps. The total preparation time is 5-6 h. The reliability of the voltage-induced release method was validated by (i) measuring the ratio of D-to L-enantiomers of the amino acids aspartic acid, alanine, and leucine, and (ii) by specific PCR amplification of four single-copy markers of human chromosome 17 and 18. We compare voltage-induced release to a frequently used silica-based protocol. DNA extracted employing voltage-induced release was more effective in PCR amplifications, which may be attributed to the effective removal of PCR inhibitors.
Bachmann
L
L
Pritzker Laboratory for Molecular Systematics and Evolution, The Field Museum, Chicago, IL 60605-2496, USA. bachmann@fmppr.fmnh.org
Scholz
M
M
Broghammer
M
M
Giddings
I
I
Pusch
C M
CM
D004247:DNA / Q000302:isolation & purification*; D004553:Electric Conductivity; D005580:Fossils; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods
D004247:DNA