芯片数据上传GEO操作指引.PDF

芯片数据上传GEO操作指南 一、登陆GEO网站下载Excel表格 1、GEO网站/geo/info/submission.html 点击箭头标注处 PS：后续以上传Agient芯片为例，供参考。 2 、下载表格 点击箭头标注处下载表格 PS：根据芯片进行选择，以“one-color experiment”为例。 二、数据整理 1、打开表格，进行“Metadata Template ”和 “Matrix Template ”填写 “Metadata Template ”填写参考“Metadata Example ”； “Matrix Template ”填写参考“Matrix Example ”； 1 2 、“Metadata Template ”填写：参考“Metadata Example ” （1）填写说明 2 （2 ）可参考的信息： growth protocol 需客户填写 treatment protocol 需客户填写 extract protocol RNA was extracted by mirVanaTM RNA Isolation Kit (Applied Biosystem p/n AM1556 ) following the manufacturers instructions. label protocol Cyanine-3 (Cy3) labeled cRNA was prepared from 0.2 μg RNA using the One-Color Low RNA Input Linear Amplification PLUS kit (Agilent) according to the manufacturers instructions, followed by RNAeasy column purification (QIAGEN, Valencia, CA). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer. hyb protocol 0.6 μg of Cy3-labelled cRNA (specific activity 10.0 pmol Cy3/μg cRNA) was fragmented at 60°C for 30 minutes in a reaction volume of 22.5 μl containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 22.5μl of 2x Agilent hybridization buffer was added to the fragmentation mixture and