生物芯片技术及其应用.pptVIP

生物芯片技术及其应用 南京医科大学第一附属医院 临床检验中心 童明庆 1.What is a microarray Microarray 或 Biochip （生物芯片）是一种微型多参数生物传感器。它通过在一微小的基片表面固定大量的分子识别探针，或构建微分析单元和系统，实现对化合物、蛋白质、核酸、细胞或其它生物组分准确、快速、大信息量的筛选或检测。 A microarray is an ordered array of microscopic elements on a planar substrate that allows the specific binding of genes or gene products. 1.What is a microarray Four distinct characteristics of microarray : Microscopic target elements or spots; Planar substrates; Rows and columns of elements; Specific binding between microarray target element on the substrate and probe molecules in solution. 1.What is a microarray Which is the probe?and which is the target? There are some differences between China and United States of America. In U.S.A. the probe are usually in solution. In China the probe is a reagent to detect target from sample. Biochips的主要类别 ?蛋白芯片 Protein chips(protein microarray)： 肽芯片，蛋白芯片 ?基因芯片 Gene chips（Gene microarray）： 靶DNA芯片，多探针芯片 ③芯片实验室Laboratory-on-a-chip or Microfluidics-based chip 2.Microarray manufacturing The three main manufacturing methods: Contact printing:uses direct contact between the printing implement or the sample contained in the implement and the microarray substrate; Noncontact printing:on-the-fly printing including ink-jet tech.,microsolenoid tech.,and thermal bubble jet dispenser; Semiconductor tech: photolithographic masks and micromirrors. Microscopic elements: ①Semiconductor tech produce 15~30μm features ② Printed spot size is 50~350 μm ③ Tissue spot is 200~600 μm Macroarrays contain sample spot sizes of about 300 microns or larger microarray are typically less than 200 microns in diameter Gene chips的制备 ? 在片原位光刻合成 O d NTP- 5′-光敏保护基因  Gene chips的制备 cDNA点样法 醛基修饰玻片 Silanized slide（与DNA上氨基反应） Silane slide 氨基修饰玻片 Silylated slide（UV照射与DNA交联） Polylysine slide 玻片氨基化举例： Nytran尼龍膜 点样前变性 變性的終濃度1×0.4N NaOH/10mM EDTA， 變性的擴增産物被點樣到N