SINGLE CELL PYROSEQUENCING USING IMPROVED LINEAR-AFTER-THE-EXPONENTIAL (ILATE) -PCR
Although the nucleic acid analysis in a single cell is very important to the research of cellular heterogeneity, the extremely small amount of template and low detection sensitivity increase the difficulty. It is requires more effective techniques that can preferably be used at the single-cell leveL Here we describe a single cell pyrosequencing assay. Using improved low-volume (as low as 1 μL) linear-afterthe-exponential-PCR on the modified glass slide combined with high sensitivity pyrosequencing, we detected the 1 555A>G mutation of inherited deafness on mitochondrial DNA. The dNTPs and single strand PCR product were treated by biotinylated PPase to reduce background noise produced by PPL The result indicated that our method has high sensitivity, simple operation and low cost, is also suitable for detecting a mass of samples. It is promising that this assay is used in the fields of diagnose of single cell.
Single cell analysis (SCA) Pyrosequencing Improved linear-after-the-exponential (imLATE)-PCR Low volume PCR Nucleic acid analysis
Song Qinxin Zhou Guohua
China Pharmaceutical University, Nanjing 210009, China Huadong Research Institute for Medicine and Biotechnics, Nanjing 210002, China Medical School, Nanji
国际会议
重庆
英文
387-389
2011-10-28（万方平台首次上网日期，不代表论文的发表时间）