High-speed analysis of multiplexed short tandem repeats with an electrophoretic microdevice.
We report the development of a robust and effective method for multiplexed short tandem repeat (STR) analysis within a chip-based microdevice. The method uses a laser-induced fluorescence detection system and simultaneously detects three- and four-color multiplexed polymerase chain reaction (PCR) samples. Analyses of the eight combined DNA index system (CODIS) STR loci were performed in 20 min with single-base-pair resolution ranging from 0.75 to 1. A simultaneous analysis of fifteen loci-ladders and a gender marker Amelogenin based on the PowerPlexTM 16 System was achieved in less than 35 min. The system is capable of repetitive operation and may be extended to high-throughput multilane devices that could be readily interfaced to an automated sample loading system.
Mitnik
Luba
L
Whitehead Institute for Biomedical Research, Cambridge, MA 02142, USA.
Carey
Loucinda
L
Burger
Robert
R
Desmarais
Samantha
S
Koutny
Lance
L
Wernet
Oliver
O
Matsudaira
Paul
P
Ehrlich
Daniel
D
D004586:Electrophoresis / Q000295:instrumentation / Q000379:methods; D008832:Microchemistry; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D020080:Tandem Repeat Sequences