Zou Z L, Wang S Q, Wang Z Q
Beijing Institute of Radiation Medicine, Beijing 100850, China.
Sheng Wu Gong Cheng Xue Bao. 2001 Sep;17(5):498-502.
The process for preparing the aldehyde slides was optimized and the properties of the aldehyde microscopic slides for immobilizing oligonucleotide were explored. The result shows that the concentration of aminosliane reagent plays an important role in the fluorescent background. Aldehyde slides with 2% aminosilane and 5% aldehyde treatment for 16 min and 30 min respectively immobilize oligonucleotide efficiently and have low fluorescence background. During oligonucleotide immobilization, terminal amino modification has no obvious specificity, but it can enhance the hybridization capacity of immobilized oligonucleotides. At low concentration (less than 10 mumol/L), hybridization signal has linear relationship with probe concentration, the hybridization signal reaches saturation when probe concentration is more than 20 mumol/L.
优化了醛基载玻片的制备工艺,并对用于固定寡核苷酸的醛基显微镜载玻片的性能进行了探索。结果表明,氨基硅烷试剂的浓度对荧光背景起着重要作用。分别用2%氨基硅烷和5%醛处理16分钟和30分钟的醛基载玻片能有效固定寡核苷酸且荧光背景较低。在寡核苷酸固定过程中,末端氨基修饰没有明显的特异性,但它可以增强固定化寡核苷酸的杂交能力。在低浓度(小于10μmol/L)时,杂交信号与探针浓度呈线性关系,当探针浓度超过20μmol/L时杂交信号达到饱和。