Tang K, Fu D J, Julien D, Braun A, Cantor C R, Köster H
Sequenom Inc., 11555 Sorrento Valley Road, San Diego, CA 92121, USA.
Proc Natl Acad Sci U S A. 1999 Aug 31;96(18):10016-20. doi: 10.1073/pnas.96.18.10016.
Silicon chips with immobilized target DNAs were used for accurate genotyping by mass spectrometry. Genomic DNAs were amplified with PCR, and the amplified products were covalently attached to chip wells via N-succinimidyl (4-iodoacetyl)aminobenzoate (SIAB) chemistry. Primer annealing, extension, and termination were performed on a 1-microl scale directly in the chip wells in parallel. Diagnostic products thus generated were detected in situ by using matrix-assisted laser desorption ionization mass spectrometry. This miniaturized method has the potential for accurate, high-throughput, low-cost identification of genetic variations.
带有固定化靶DNA的硅芯片被用于通过质谱进行精确的基因分型。基因组DNA通过PCR进行扩增，扩增产物通过N-琥珀酰亚胺基（4-碘乙酰基）氨基苯甲酸酯（SIAB）化学方法共价连接到芯片孔中。引物退火、延伸和终止在芯片孔中直接以1微升规模并行进行。由此产生的诊断产物通过基质辅助激光解吸电离质谱进行原位检测。这种小型化方法具有精确、高通量、低成本鉴定基因变异的潜力。
