Low volume amplification and sequencing of mitochondrial DNA on a chemically structured chip.
Low volume (LV) amplification (1 microL) of nuclear DNA (nucDNA) on a chemically structured chip is an appropriate and highly sensitive method to simultaneously amplify amelogenin and 15 forensically relevant short tandem repeats (STR). In this study, a combined method using on-chip LV amplification of mitochondrial DNA (mtDNA) and subsequent on-chip LV cycle sequencing was established to obtain a method, which is sensitive and robust enough to allow reliable analysis of DNA amounts representing the single cell level. All the necessary steps of the procedure--except for the purification of the sequencing products--were accomplished within the same final 2-microL reaction volume.
Lutz-Bonengel
Sabine
S
Institute of Legal Medicine, Albert Ludwig University Freiburg, Albertstrasse 9, 79104, Freiburg, Germany. sabine.lutz-bonengel@uniklinik-freiburg.de
Sänger
Timo
T
Heinrich
Marielle
M
Schön
Ulrike
U
Schmidt
Ulrike
U
D016172:DNA Fingerprinting / Q000379:methods*; D004272:DNA, Mitochondrial / Q000032:analysis* / Q000235:genetics; D046209:Electrophoresis, Microchip / Q000379:methods*; D006801:Humans; D018895:Microsatellite Repeats / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*
D004272:DNA, Mitochondrial