摘要
将巯基修饰的寡核苷酸探针通过自组装技术固定于以光刻溅射技术制作的石英金片上来制备核酸传感器探头。然后将探头与生物素标记的21个碱基目标寡核苷酸杂交，再利用生物素－亲合素的作用联入亲合素标记的碱性磷酸酯酶，酶催化底物AMPPD发光来达到测定寡核苷酸的目的。测得杂交清洗温度70℃时探头对错配碱基核酸的选择性最佳，探头的测定范围为0.1～100靏/L，最低检测浓度为0.1靏/L，重复测定的相对标准偏差为11.3%。
A nucleic acid biosensor based on self-assembled monolayers technology was developed, thiolated oligonucleotide was immobilized on gold film which deposed on quartz slides by photolithographic technique. Immobilized oligonucleotide was hybridized with biotinylated target 'oligonucleotide. Then extravidin-alkaline phosphatase which can dephosphorylate the substrates adamantyl-1,2-dioxetane phosphate (AMPPD) to produce a measureable yield of fight output was bound with biotinylated target oligonucleotide. An optimum selectivity for sequence mismatch target oligonucleotide was obtained at 70 degreesC washing temperature after hybridization. The target detection range of 0.1 similar to 100 mug/L, sensitivity of 0.1 mug/L were obtained, relative standard drivation was 11.3% for the determination of 100 mug/L target (n = 10).
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
2001年第9期997-999,共3页
Chinese Journal of Analytical Chemistry
基金
国家自然科学基金资助项目(No.29805005)
