TY - JOUR
T1 - Development and evaluation of an automated workstation for single nucleotide polymorphism discrimination using bacterial magnetic particles
AU - Tanaka, Tsuyoshi
AU - Maruyama, Kohei
AU - Yoda, Kiyoushi
AU - Nemoto, Etsuo
AU - Udagawa, Yuuji
AU - Nakayama, Hideki
AU - Takeyama, Haruko
AU - Matsunaga, Tadashi
N1 - Funding Information:
This work was funded in part by a Grant-in-Aid for Specially Promoted Research, No. 13002005 and Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology. It was also supported by NEDO's Subsidy for FY2002 University-Initiated Practical Business Creation R&D Program.
PY - 2003/12/15
Y1 - 2003/12/15
N2 - We designed an automated workstation for magnetic particle-based single nucleotide polymorphism (SNP) discrimination of ALDH genotypes. Bacterial magnetic particles (BMPs) extracted from Magnetospirillum magneticum AMB-1 were used as DNA carriers. The principle for SNP discrimination in this study was based on fluorescence resonance energy transfer (FRET) between FITC (donor) and POPO-3 (acceptor) bound to double-stranded DNA. The workstation is equipped with a 96-way automated pipetter which collects and dispenses fluids as it moves in x- and z-directions. The platform contains a disposable tip rack station, a reagent vessel serving as a stock for POPO-3 and FITC-labeled probes and a reaction station for a 96-well microtiter plate. BMPs were collected by attaching a neodymium iron boron sintered (Nd-Fe-B) magnet on the bottom of the microtiter plate. This system permits the simultaneous heating and magnetic separation of 96 samples per assay. The genotypes ALDH2*1 and ALDH2*2 were discriminated by calculating the relative fluorescence intensities on BMPs.
AB - We designed an automated workstation for magnetic particle-based single nucleotide polymorphism (SNP) discrimination of ALDH genotypes. Bacterial magnetic particles (BMPs) extracted from Magnetospirillum magneticum AMB-1 were used as DNA carriers. The principle for SNP discrimination in this study was based on fluorescence resonance energy transfer (FRET) between FITC (donor) and POPO-3 (acceptor) bound to double-stranded DNA. The workstation is equipped with a 96-way automated pipetter which collects and dispenses fluids as it moves in x- and z-directions. The platform contains a disposable tip rack station, a reagent vessel serving as a stock for POPO-3 and FITC-labeled probes and a reaction station for a 96-well microtiter plate. BMPs were collected by attaching a neodymium iron boron sintered (Nd-Fe-B) magnet on the bottom of the microtiter plate. This system permits the simultaneous heating and magnetic separation of 96 samples per assay. The genotypes ALDH2*1 and ALDH2*2 were discriminated by calculating the relative fluorescence intensities on BMPs.
KW - Aldehyde dehydrogenase 2 gene
KW - Bacterial magnetic particles (BMPs)
KW - Fluorescence resonance energy transfer
KW - Single nucleotide polymorphism (SNP)
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U2 - 10.1016/S0956-5663(03)00189-1
DO - 10.1016/S0956-5663(03)00189-1
M3 - Article
C2 - 14615090
AN - SCOPUS:0242570444
SN - 0956-5663
VL - 19
SP - 325
EP - 330
JO - Biosensors and Bioelectronics
JF - Biosensors and Bioelectronics
IS - 4
ER -