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1、 等位基因鉴别实验 美国应用生物系统公司 7300 7500 实时定量 PCR 仪 简介和示例实验简介和示例实验 设定反应板 设计等位基因 鉴别实验 读取等位基因鉴 别实验扩增前 信号 生成 扩增数据 设定反应板 设计等位基因 鉴别实验 读取等位基因鉴 别实验扩增前 信号 生成 扩增数据 读取等位基因鉴 别实验扩增后 信号 读取等位基因鉴 别实验扩增后 信号 3 30 2004 8 25 AD GSG FrontCover fm ii7300 7500 实时定量 PCR 仪等位基因鉴别实验入门指南 Copyright 2004 Applied Biosystems All rights res
2、erved For Internal Use Only For Research Use Only Not for use in diagnostic procedures Authorized Thermal Cycler This instrument Serial No is an Authorized Thermal Cycler Its purchase price includes the up front fee component of a license under United States Patent Nos 4 683 195 4 683 202 and 4 965
3、188 owned by Roche Molecular Systems Inc and under corresponding claims in patents outside the United States owned by F Hoffmann La Roche Ltd covering the Polymerase Chain Reaction PCR process to practice the PCR process for internal research and development using this instrument The running royalty
4、 component of that license may be purchased from Applied Biosystems or obtained by purchasing Authorized Reagents This instrument is also an Authorized Thermal Cycler for use with applications licenses available from Applied Biosystems Its use with Authorized Reagents also provides a limited PCR lic
5、ense in accordance with the label rights accompanying such reagents Purchase of this product does not itself convey to the purchaser a complete license or right to perform the PCR process Further information on purchasing licenses to practice the PCR process may be obtained by contacting the Directo
6、r of Licensing at Applied Biosystems 850 Lincoln Centre Drive Foster City California 94404 DISCLAIMER OF LICENSE No rights for any application including any in vitro diagnostic application are conveyed expressly by implication or by estoppel under any patent or patent applications claiming homogeneo
7、us or real time detection methods including patents covering such methods used in conjunction with the PCR process or other amplification processes The 5 nuclease detection assay and certain other homogeneous or real time amplification and detection methods are covered by United States Patent Nos 5
8、210 015 5 487 972 5 804 375 and 5 994 056 owned by Roche Molecular Systems Inc by corresponding patents and patent applications outside the United States owned by F Hoffmann La Roche Ltd and by United States Patent Nos 5 538 848 and 6 030 787 and corresponding patents and patent applications outside
9、 the United States owned by Applera Corporation Purchase of this instrument conveys no license or right under the foregoing patents Use of these and other patented processes in conjunction with the PCR process requires a license For information on obtaining licenses contact the Director of Licensing
10、 at Applied Biosystems 850 Lincoln Centre Drive Foster City California 94404 or The Licensing Department Roche Molecular Systems Inc 1145 Atlantic Avenue Alameda California 94501 USA Information in this document is subject to change without notice Applied Biosystems assumes no responsibility for any
11、 errors that may appear in this document This document is believed to be complete and accurate at the time of publication In no event shall Applied Biosystems be liable for incidental special multiple or consequential damages in connection with or arising from the use of this document TRADEMARKS App
12、lied Biosystems MicroAmp ROX and VIC are registered trademarks of Applera Corporation or its subsidiaries in the U S and or certain other countries AB Design Applera ABI PRISM Assays by Design Assays on Demand BloodPrep Celera Genomics FAM iScience iScience Design and NucPrep are trademarks of Apple
13、ra Corporation or its subsidiaries in the U S and or certain other countries SYBR Green is a registered trademark of Molecular Probes Inc AmpliTaq Gold AmpErase and TaqMan are registered trademarks of Roche Molecular Systems Inc All other trademarks are the sole property of their respective owners P
14、art Number 4347964 Rev B 3 2004 7300 7500 PCR iii TaqMan 89 56 A BC DE56F GH56IJ 34 56K 89 3456K 89 LMNO 1 2 3 4 5 6 7300 7500 PCR A BC PQRS TU BC iv7300 7500 PCR 目录 SAMPLE DOCUMENT March 30 2004 8 25 pm AD GSG TOC fm 7300 7500 实时定量 PCR 仪等位基因鉴别实验入门指南v 等位基因鉴别实验工作流程等位基因鉴别实验工作流程iii 前言前言vii 第第 1 章简介和等位基
15、因鉴别示例实验章简介和等位基因鉴别示例实验1 概述 1 关于 7300 7500 PCR 仪 2 关于等位基因鉴别 AD 分析 2 关于等位基因鉴别实验 4 等位基因鉴别示例实验 6 第第 2 章设计等位基因鉴别实验章设计等位基因鉴别实验13 工作流程 13 使用 TaqMan 探针法的序列检测化学试剂 14 选择探针和引物 15 第第 3 章设定反应板章设定反应板17 工作流程 17 准备 DNA 18 设定反应板 19 第第 4 章读取等位基因鉴别实验扩增前信号章读取等位基因鉴别实验扩增前信号23 工作流程 23 扩增前荧光信号读取 24 开始之前 24 创建等位基因鉴别反应板文件 24
16、读取扩增前荧光信号 29 第第 5 章生成扩增数据章生成扩增数据31 工作流程 31 创建绝对定量 AQ 反应板文件 32 执行扩增程序 36 SAMPLE DOCUMENT March 30 2004 8 25 pm AD GSG TOC fm vi7300 7500 实时定量 PCR 仪等位基因鉴别实验入门指南 第第 6 章读取和分析等位基因鉴别扩增后信号章读取和分析等位基因鉴别扩增后信号39 工作流程 39 读取扩增后荧光信号 40 评估结果 42 等位基因分型 43 查看报告 45 导出反应板文件 46 附录附录 A创建探针创建探针47 附录附录 B查看扩增数据查看扩增数据49 设置分析参数 49 分析扩增数据 绝对定量反应板 50 查看扩增数据 51 参考文献参考文献55 索引索引57 DRAFT 3 30 2004 8 25 AD GSG Preface fm 7300 7500 实时定量 PCR 仪等位基因鉴别实验入门指南vii 前言 如何使用本指南 本指南的目的本指南为使用美国应用生物系统公司 7300 7500 实时定量 PCR 仪 7300 7500 PCR 仪 进
