医疗器械-软件设计和开发-验证报告-模板范例

医疗器械软件设计开发验证报告医疗器械软件设计开发验证报告PagePage#of70医疗器械软件设计开发验证报告记录编号：目ODD录DTOC\o"1-5"\h\z\o"CurrentDocument"\h第一部分总则 2\o"CurrentDocument"\h一、概述 2...\o"CurrentDocument"\h二、验证方案 2.1、验证目的 2.2、验证范围 2.\o"CurrentDocument"\h3、相关文件 2.4、职责 2.5、验证条件 3.6、验证方法 3.7、验证合格标准及评价分析 7.8、验证实施计划 7.9、验证周期 8.10、数据收集 8.\o"CurrentDocument"\h第二部分测试确认 8\o"CurrentDocument"\h一、功能性测试： 8.\o"CurrentDocument"\h二、性能测试： 9.\o"CurrentDocument"\h三、可靠性和安全性测试 9\o"CurrentDocument"\h四、用户场景测试 9.\o"CurrentDocument"\h五、准确性测试 1. 0普通碱基测试 10\o"CurrentDocument"\h修饰碱基测试 16\o"CurrentDocument"\h兼并碱基测试 53\o"CurrentDocument"\h含RNA碱基测试 59含mRNA碱基测试 63\o"CurrentDocument"\h含特殊碱基I序列测试 65含特殊碱基U序列测试 67\o"CurrentDocument"\h第三部分结论 70第一部分总则一、概述本次验证严格按照验证方案要求实施，整个过程稳定可控，验证过程没有出现偏差，Filemaker的各项性能符合相关规定。二、验证方案1、验证目的通过验证以证明使用Filemaker软件设计的Filemaker数据库系统在适用性、准确性、稳定性、可控性、安全性和保密性方面可以满足公司质量管理体系和用户的要求与需要。2、验证范围使用Filemaker软件设计的以下功能模块：Filemaker.fp73、相关文件文件名称文件描述DP_U_0025Filemaker操作方法QP_U_0114文件与资料的保密管理DP_U_0140Filemaker数据库应急方案与备份方案DP_U_0117Filemaker账户权限说明4、职责验证委员会负责验证方案的审批；负责验证的协调工作，以保证本验证方案规定项目的顺利实施；负责验证数据及结果的审核；负责验证报告的审批。验证委员会名单：姓名部门职位职责验证小组负责验证方案的起草、修改；负责组织本验证方案的实施；负责验证数据的统计、分析、审核，报验证委员会审核；负责验证报告的编写，并报验证委员会。验证小组名单：5、验证条件FilemakerServerAdvance9.2,数据库托管服务器端；WindowsServer2003,数据库托管服务器端操作系统;FilemakerPro11.0,数据库客户端；WindowsXP,数据库客户端操作系统；InternetExplorer7.0,WebPublishing访问客户端。6、验证方法质量策划风险管理计划见：《QR_U_0041_Filemaker软件验证报告》配制管理计划见：《QR_U_0041_Filemaker软件验证报告》系统需求定义见《Filemaker数据库需求分析》设计文档见《Filemaker总体设计说明书》测试方法

功能性测试对FilemakerFilemaker数据库的各项功能进行测试，以验证其适合性、互操作性和保密与安全性符合QMS的要求：特性测试内容说明符合要求判定适合性文件的共享与托管，即时网络发布使用FSA托管并发布数据库，使用FilemakerPro和IE两种方式进行访问查找所需数据分别测试一般搜索、限制搜索和扩展搜索功能，导出搜索结果数据的导入导出批量导入，导出文件批量导入，导出文件信息数据库布局的创建与设计根据需要更改布局、字段、脚本保密与安全性账户和权限集管理员可以控制增加任意数量的账户。测试系统及其数据访问的可控制性。测试系统防止非法操作的模式，包括防止非授权的创建、删除和修改程序或信息可控的记录审核流程可增加或修改审核流程，可查看审核状态自动备份可设置自动备份周期，自动备份数据库限制编辑测试系统防止数据被误删、误改和被破坏的能力测试者：年月日验证小组确认人签字：年月日性能测试对Filemaker构建的Filemaker数据库系统的性能进行强度测试，以验证其系统响应时间、系统存储量和异常状态系统性能符合QMS的要求：测试系统的响应时间，包括单个用户、多用户并发的情况，批量导入记录的响应时间；测试系统的最大存储量、100条记录的平均存储量；测试运行条件在异常状态下，或在人为设定的状态下，系统的性能。

特性测试内容响应时间系统响应时间单客户端访问数据库单IE浏览器访问数据库5个客户端同时访问数据库10个IE浏览器同时访问数据库批量导入100条记录的响应时间系统存储量数据库最大存储量100条记录平均存储量异常状态系统性能服务器重启后系统性能系统恢复后的性能域环境杀毒软件扫描阶段系统的性能测试者： 验证小组确认人签字：年月日 年月日可靠性和安全性测试见：《QR_U_0041_Filemaker软件验证报告》用户场景测试测试Filemaker.fp7的实际应用场景，以用户角色权限完成一项特定的业务处理流程测试其易理解性、易学性、易操作性和时间特性：特性测试内容说明符合要求判定易理解性系统的各项功能是否容易被识别或被理解参考Filemakerintroduction-databasesolutionQR_U_0001_数据库培训20100902界面的输入和输出的格式和含义是否容易被理解易学性系统的在线帮助是否容易定位，是否有效各数据库负责人和管理员是否明确及联络方式是否有效测试用户文档的有效性测试“3、相关文件”中所有文件的有效性易操作性系统是否对输入数据进行有效性检查以正确操作、误操作模式、非常规操作

中断执行功能是否可以在动作完成之前被取消模式为框架设计测试用例。误操作模式有错误的数据类型作参数、错误的输入数据序列、错误的操作序列。确认参数是否易于选择，是否有缺省值解释的消息是否明确界面提示是否有效确认系统能否提示出错的风险、能否纠正错误的输入、能否从错误中恢复确认定制能力的有效性时间特性系统响应时间完成一项规定的任务所需的时间平均响应时间执行若干并行任务所需的平均时间响应极限时间最大负载条件下，系统完成某项任务需要时间的极限系统的吞吐量给定的时间周期内系统能完成的任务数量平均吞吐量在一个单位时间内系统处理并发任务的平均数极限吞吐量最大负载条件下，在给定的时间周期内系统处理的最多并发任务数测试者：年月日验证小组确认人签字：年月日准确性测试随机抽取100条普通碱基序列，验证普通碱基的分子量、TM值和消光系数计算与IDT网站的差异率。所有兼并碱基对随机组合，每条序列最多三对，不包含兼并碱基对位于序列首位的情况，验证兼并碱基的分子量、TM值和消光系数计算与IDT网站的差异率。所有修饰碱基随机组合，每条序列最多四种，序列中不包含兼并碱基、特殊碱基和IDT网站上没有的修饰碱基，验证修饰碱基的分子量、TM值和消光系数计算与IDT网站的差异率。随机选取以下四种含RNA的序列个10条，验证含RNA的序列的分子量、TM值和消光系数计算与IDT网站的差异率。全部为RNA5′端部分含RNA中间部分含RNA3′端部分含RNA随机选取以下三种含mRNA的序列各5条，验证含mRNA的序列的分子量、TM值和消光系数计算与IDT网站的差异率。5′端部分含RNA中间部分含RNA3′端部分含RNA6.5.5.5随机选取以下三种含特殊碱基I的序列各5条，验证含I的序列的TM值和消光系数计算与IDT网站的差异率。5′端部分含I中间部分含I3′端部分含I6.5.5.5随机选取以下三种含特殊碱基U的序列各5条，验证含U的序列的TM值和消光系数计算与IDT网站的差异率。5′端部分含U中间部分含U3′端部分含U7、验证合格标准及评价分析Filemaker.fp7数据库的适合性、互操作性和保密与安全性符合QMS的要求；Filemaker.fp7数据库的系统响应时间、系统存储量和异常状态系统性能应符合用户的要求；Filemaker.fp7数据库的可靠性和安全性应符合QMS的要求；Filemaker.fp7数据库的易理解性、易学性、易操作性和时间特性应能够适应用户的要求；Filemaker.fp7数据库的应急方案应能保证其安全性和可持续性。Filemaker.fp7数据库的准确性应符合用户要求。8、验证实施计划验证步骤时间安排Filemaker.fp7导入2019-9-6准确性验证Filemaker.fp7试用2019-9-13到2019-9-31

9、验证周期10、数据收集第二部分测试确认、功能性测试：特性测试内容说明符合要求判定适合性文件的共享与托管，即时网络发布使用FSA托管并发布数据库，使用FilemakerPro和IE两种方式进行访问查找所需数据分别测试一般搜索、限制搜索和扩展搜索功能，导出搜索结果数据的导入导出批量导入，导出文件批量导入，导出文件信息数据库布局的创建与设计根据需要更改布局、字段、脚本保密与安全性账户和权限集管理员可以控制增加任意数量的账户。测试系统及其数据访问的可控制性。测试系统防止非法操作的模式，包括防止非授权的创建、删除和修改程序或信息可控的记录审核流程可增加或修改审核流程，可查看审核状态自动备份可设置自动备份周期，自动备份数据库限制编辑测试系统防止数据被误删、误改和被破坏的能力测试者：年月日验证小组确认人签字：年月日

二、性能测试：特性测试内容响应时间系统响应时间单客户端访问数据库单IE浏览器访问数据库5个客户端同时访问数据库10个IE浏览器同时访问数据库批量导入100条记录的响应时间系统存储量数据库最大存储量100条记录平均存储量异常状态系统性能服务器重启后系统性能系统恢复后的性能域环境杀毒软件扫描阶段系统的性能测试者： 验证小组确认人签字：年月日 年月日三、可靠性和安全性测试见：《QR_U_0041_Filemaker软件验证报告》四、用户场景测试特性测试内容说明符合要求判定易理解性系统的各项功能是否容易被识别或被理解参考Filemakerintroduction-databasesolutionQR_U_0001_数据库培训20100902界面的输入和输出的格式和含义是否容易被理解易学性系统的在线帮助是否容易定位，是否有效各数据库负责人和管理员是否明确及联络方式是否有效测试用户文档的有效性测试“3、相关文件”中所有文件的有效性易操作性系统是否对输入数据进行有效性检查以正确操作、误操作模式、非常规操作模式为框架设计测试用例。误操作模式有错误的数据类型作参数、错误的输入中断执行功能是否可以在动作完成之前被取消

特性测试内容说明符合要求判定确认参数是否易于选择，是否有缺省值数据序列、错误的操作序列。解释的消息是否明确界面提示是否有效确认系统能否提示出错的风险、能否纠正错误的输入、能否从错误中恢复确认定制能力的有效性时间特性系统响应时间完成一项规定的任务所需的时间平均响应时间执行若干并行任务所需的平均时间响应极限时间最大负载条件下，系统完成某项任务需要时间的极限系统的吞吐量给定的时间周期内系统能完成的任务数量平均吞吐量在一个单位时间内系统处理并发任务的平均数极限吞吐量最大负载条件下，在给定的时间周期内系统处理的最多并发任务数测试者：验证小组确认人签字：年月日年月日五、准确性测试1普通碱基测试1.1随机抽取100条普通碱基序列，验证普通碱基的分子量、TM值和消光系数计算与IDT网站的差异率。NO碱基序列FM-MWFM-TM(℃)FM-εIDT-MWIDT-TM(℃)IDT-εMW差值TM差值ε差值15'-CACCAAAACTTTCTCTCTTgATA-3'6917.650.72139006917.650.72139000.00.0025'-TgAgAAAggCATTAgAAAgC-3'6223.150.32128006223.150.42128000.00.1035'-CAgCCACCAgCACATCAAT-3'5710.855.71819005710.855.91819000.00.20

NO碱基序列FM-MWFM-TM(℃)FM-εIDT-MWIDT-TM(℃)IDT-εMW差值TM差值ε差值45'-gTTTTACTTACTCTCgTCTCCACAg-3'7533.954.82239007533.954.92239000.00.1055'-CAATTACTTCggCTTTAAgAC-3'6380.249.61992006380.249.31992000.0-0.3065'-TTTCAgCCACTTTTCCATT-3'5679.750.21648005679.7501648000.0-0.2075'-AgAgACAAAAAgggAgTAACT-3'6545.351.22315006545.3512315000.0-0.2085'-CAgTgCCACTggTCTATAAT-3'6092.051.91907006092.051.71907000.0-0.2095'-gCCATTCTggTggACCT-3'5177.455.01532005177.454.71532000.0-0.30105'-CCCCACTCTgACCAAAg-3'5084.452.81561005084.452.51561000.0-0.30115'-TCCTgACTgTCATCCCTATTgg-3'6652.455.61966006652.455.91966000.00.30125'-ATCTgggAAgAACTCAgCgAAC-3'6777.556.62223006777.556.92223000.00.30135'-ggAggAgTCAggggCAgAAT-3'6296.158.72106006296.158.92106000.00.20145'-CACAggATggggAAgTCACAg-3'6529.357.82165006529.357.62165000.0-0.20155'-CAAgCATgCAggAAgAggAg-3'6249.155.92120006249.155.92120000.00.00165'-ggTgCCTAgCTTgTATgTTg-3'6170.053.31859006170.053.61859000.00.30175'-gCgAgCCTgTCAAgTCATAg-3'6142.055.51958006142.055.51958000.00.00185'-ACTgTCCCAAAATCgTCTAT-3'6036.050.91909006036.051.11909000.00.20195'-TCTCCTAggCTCAAgCAgTg-3'6093.056.31860006093.056.31860000.00.00205'-CCgCATTAAAgCTACCACTA-3'6030.052.41926006030.052.21926000.0-0.20215'-gTTgCCAATgATgTTACAgA-3'6156.150.52008006156.150.72008000.00.20225'-gCTggATgATggggCgATTC-3'6229.158.51923006229.158.71923000.00.20235'-gAAgCATACATCCCgAAAgA-3'6128.152.42093006128.152.22093000.0-0.20245'-CTgCTTggTgTTggCgTTgA-3'6186.058.61810006186.058.71810000.00.10255'-CTCCATgACCTgCgCCTgTg-3'6044.961.01731006044.9611731000.00.00265'-AAggCCgAggTgATggTggA-3'6287.160.82073006287.161.22073000.00.40275'-ATTCTTTAgAggTgTTggAT-3'6193.148.71970006193.148.51970000.0-0.20285'-AACTCCCAATCTATAgCCAg-3'6030.051.41945006030.051.21945000.0-0.20295'-TgCCTACAgTAAgCTCTATg-3'6092.051.31904006092.051.11904000.0-0.20305'-AATggCCgCTTTTTgTgCTAA-3'6427.255.91951006427.256.11951000.00.20315'-ACACAATAAACTTCACCAAA-36031.047.52065006031.047.62065000.00.10

NO碱基序列FM-MWFM-TM(℃)FM-εIDT-MWIDT-TM(℃)IDT-εMW差值TM差值ε差值325'-AgATTTCTggCACTTTATTA-3'6097.046.41926006097.046.51926000.00.10335'-CCTggTgAgTTTgggATT-3'5576.751.81693005576.7521693000.00.20345'-CTgATgAgACAgCAACAA-3'5525.749.91866005525.749.61866000.0-0.30355'-CAgggTgATAAAAgAgCAAT-3'6223.150.12138006223.150.22138000.00.10365'-CAAAACTggAACAATCAgCC-3'6088.052.42001006088.052.32001000.0-0.10375'-TgCgTgTACTTCggCTgACT-3'6115.058.81791006115.058.91791000.00.10385'-gggCgAACgACAggAAgATg-3'6265.158.12102006265.158.32102000.00.20395'-CgCATCgTCATCggCTCCAA-3'6038.060.01832006038.060.31832000.00.30405'-CAAACggCgTATCAATTCAC-3'6070.052.41949006070.052.31949000.0-0.10415'-CTCTggTACgAACgCCTCAC-3'6038.057.51808006038.057.61808000.00.10425'-AgCCCTTgTgACTCCAgCAg-3'6078.059.51854006078.059.71854000.00.20435'-CCgCAgCgACTTCCACgATg-3'6063.061.21831006063.061.21831000.00.00445'-TgTgTgATggCgTTgAAgAT-3'6243.154.71981006243.154.71981000.00.00455'-CTgTTggCATTCgggAAAgT-3'6188.155.41920006188.155.81920000.00.40465'-gTgCAAAggTAgCATAATCA-3'6174.150.32094006174.150.52094000.00.20475'-TgTCCTgATCCAACATCgAg-3'6077.053.91903006077.054.21903000.00.30485'-CggCgACgCATCTTTCAAATg-3'6406.257.21967006406.257.41967000.00.20495'-CTCACCgACgATATgCTCCg-3'6038.057.41829006038.057.51829000.00.10505'-ACTggTCgAgCTCCAATTCATgT-3'7014.658.72167007014.658.52167000.0-0.20515'-TCCCTggTgTgATCCTgCCAgT-3'6693.462.91955006693.462.61955000.0-0.30525'-gATCCTTCCTgCAggTTCACCT-3'6637.358.91943006637.3591943000.00.10535'-AAgAgCgACCggCgATgTgT-3'6207.161.41998006207.161.81998000.00.40545'-AAACTAggATTAgATACCCTATTAT-3'7648.148.72570007648.148.62570000.0-0.10555'-ACCACATATTTTCTAATgCTC-3'6315.248.21967006315.248.11967000.0-0.10565'-AATgACTgTgTTTCTTATgAC-3'6426.248.32015006426.248.22015000.0-0.10575'-CCTTCCTTCTTTTTgATTTTgTTT-3'7222.751.11995007222.750.71995000.0-0.40

NO碱基序列FM-MWFM-TM(℃)FM-εIDT-MWIDT-TM(℃)IDT-εMW差值TM差值ε差值585'-AgACTATCAACTTAATTTCTgATC-3'7285.848.62339007285.848.42339000.0-0.20595'-CCCACCTTCCCCTCTCTCCAggCAAATggg-3'9023.968.32620009023.968.22620000.0-0.10605'-gggCCTCAgTCCCAACATggCTAAgAggTg-3'9257.066.52897009257.066.62897000.00.10615'-TgTTACTAgATCgCAgATCCTCT-3'6989.654.62158006989.654.72158000.00.10625'-CTAgAAggCAATTCTAATTgATC-3'7046.649.22287007046.649.22287000.00.00635'-ACACCCAACCACCgAAACAC-3'5978.058.21945005978.058.21945000.00.00645'-AACCACCgAAACACCCAgC-3'5704.858.61848005704.858.51848000.0-0.10655'-CCCAACAACAACCACCgTTCA-3'6273.158.11991006273.158.41991000.00.30665'-TggCCCTggACgAgAgTACCT-3'6447.261.91985006447.262.11985000.00.20675'-gTCgCgggAACAgAggTgT-3'5933.959.91906005933.9601906000.00.10685'-ggTgTTCCTCCATTgCgAAA-3'6108.055.41882006108.055.81882000.00.40695'-TCCTTCCgTTTCCTCgCC-3'5328.557.41432005328.557.51432000.00.10705'-TTCCACgCCCTCTCCgCT-3'5322.561.91445005322.561.91445000.00.00715'-CCTTCATgTTCggCggTCTCg-3'6380.260.41830006380.260.51830000.00.10725'-gCgTCATgATCggCTCgATg-3'6149.058.41884006149.058.61884000.00.20735'-gACTTgCgTgTTCgTTCTTC-3'6081.053.91749006081.054.31749000.00.40745'-AACACCgTTgAgCTTgAgAC-3'6126.055.01959006126.055.31959000.00.30755'-AggTgATggTggCTgTTATg-3'6259.154.51975006259.154.91975000.00.40765'-ATgggAgTTCCTTCTTggTT-3'6145.053.81842006145.053.71842000.0-0.10775'-gACCTCCATATTACTgAAAggAAg-3'7369.952.62434007369.952.82434000.00.20785'-gAgTAATTgCTCCATCCTgTTCA-3'6989.654.72163006989.654.82163000.00.10795'-gATCTCCAgTgTgTggACCTCAA-3'7039.658.32203007039.658.52203000.00.20

NO碱基序列FM-MWFM-TM(℃)FM-εIDT-MWIDT-TM(℃)IDT-εMW差值TM差值ε差值805'-TggAgCACACCATCACACATC-3'6344.257.32015006344.257.52015000.00.20815'-TTTggAgTAgCAgCCAAAgATCAAg-3'7739.157.22556007739.157.42556000.00.20825'-ACCATCggAATCCACggAAg-3'6120.057.12012006120.057.12012000.00.00835'-CTCgTgTgTTTCAAATggAgTgg-3'7125.756.62209007125.756.42209000.0-0.20845'-TggTgACATCTgggATATTAAgTgg-3'7792.156.22509007792.156.32509000.00.10855'-gAggAggAAAAgACCgAAAAgAA-3'7221.854.82599007221.854.92599000.00.10865'-TggTggCTggggATTgTAg-3'5970.957.31848005970.957.11848000.0-0.20875'-gCCTCgCCTTgCTCgTCTAC-3'5995.960.61665005995.960.51665000.0-0.10885'-ATCgCATCAggggCACACAg-3'6136.060.01981006136.060.31981000.00.30895'-TggACCTgTggAACTgggAT-3'6213.158.21937006213.158.31937000.00.10905'-ggACCACggAAgAgCCTgA-3'5871.959.31923005871.959.31923000.00.00915'-CgCTTTgACgCAgAgTTCAC-3'6093.056.81850006093.056.81850000.00.00925'-TTggAgCAATTgTgggACAAC-3'6510.356.32083006510.356.12083000.0-0.20935'-TCCgTggCAACAACTTTgg-3'5803.856.01766005803.856.21766000.00.20945'-TTCgACTTTgCCAgCCTgTAC-3'6348.257.41858006348.257.61858000.00.20955'-CAgggAgAgCgTgCTAAg-3'5613.755.31833005613.755.31833000.00.00965'-CgTAAACgCCgTAAACgTATTCC-3'6992.656.52225006992.656.32225000.0-0.20975'-CAgCTCAgAggAggAggATgA-3'6569.358.02209006569.357.82209000.0-0.20985'-CgTAgAgTCACACTTgCAACAAAA-3'7338.855.52430007338.855.92430000.00.40995'-CCAgACgTCggCTgCTACA-3'5773.859.61772005773.859.71772000.00.101005'-CACCgAgAAgTgggTTgACA-3'6191.157.22026006191.157.22026000.00.00验证者：年月日验证小组确认人签字：年月日普通碱基分子量、Tm值和摩尔消光系数计算数据分析：通过与IDT网站计算数据的差值分析FM数据库计算的准确性：

差值分子量Tm值摩尔消光系数最大值00.40最小值0-0.40平均值00.10標準偏差00.1946840样本数100100100設定量000下限規格0-0.50上限規格00.60做TM值差值的I-MR控制图：UCL=0.556X=0.0521 11 2131UCL=0.556X=0.0521 11 213171 81 9141 51 61观测值718191111213141 51 718191111213141 51 61观测值结论：AFM数据库普通碱基序列的分子量计算与IDT网站平均差值为0。AFM数据库普通碱基序列的TM值计算与IDT网站平均差值为0.1℃，上限规格为0.6℃，下限规格为-0.5℃。AFM数据库普通碱基序列的摩尔消光系数计算与IDT网站平均差值为0。

2修饰碱基测试所有修饰碱基随机组合，每条序列最多四种，序列中不包含兼并碱基、特殊碱基和IDT网站上没有的修饰碱基，验证修饰碱基的分子量、TM值和消光系数计算与IDT网站的差异率。NO碱基序列FM-MWFM-TM(℃)FM-εIDT-MWIDT-TM(℃)IDT-εMW差值TM差值ε差值15'-(2'-F-A)AAACACCggA(2'-F-A)GTTCTCgTA(Cy5)ACCgATCT(2'-F-A)-3'9755.661.33105009755.658.93105000.0-2.4025'-(2'-F-C)AAACACCggA(2'-F-C)GTTCTCgTG(Cy5)GCCgATCT(2'-F-C)-3'9715.565.22857009715.564.72857000.0-0.5035'-(2'-F-G)AAACACCggA(2'-F-G)GTTCTCgTC(Cy5)CCCgATCT(2'-F-G)-3'9755.564.72913009755.565.52913000.00.8045'-(2'-F-U)AAACACCggA(2'-F-U)GTTCTCgTT(Cy5)TCCgATCT(2'-F-U)-3'9668.561.72885009668.558.42885000.0-3.3055'-(2,6-Diaminopurine)AAACACCggA(2,6-Diaminopurine)GTTCTCgTA(2-aminopurine)GCCgATCT(2,6-Diaminopurine)-3'9632.462.62781009632.462.22781000.0-0.4065'-(NH2C6dT)AAACACCggA(5-Br-dU)GTTCTCgTG(2-aminopurine)CCCgATCT(5-MedC)-3'9664.365.22690009664.364.42690000.0-0.8075'-(5-Br-dU)AAACACCggA(5-Me-dC)GTTCTCgTC(2-aminopurine)TCCgATCT(5-OctadiynyldU)-3'9575.162.42688009575.163.52688000.01.1085'-(5-Me-dC)AAACACCggA(5-OctadiynyldU)GTTCTCgTT(2-aminopurine)A9230.161.32773009230.161.72773000.00.40

NO碱基序列FM-MWFM-TM(℃)FM-εIDT-MWIDT-TM(℃)IDT-εMW差值TM差值ε差值CCgATCT(3'NH2C6-dT)-3'95'-(BiotindT)AAACACCggA(5-TAMRATMAzide)GTTCTCgTA(2-aminopurine)CCCgATCT(BiotindT)-3'9887.062.53106009887.062.13106000.0-0.40105'-(2'-F-A)GAACACCggA(6-FAMTMAzide)GTTCTCgTA(2-aminopurine)TCCgATCT(DABCYL)-3'9342.261.03061839342.261.43061830.00.40115'-(2'-F-C)GAACACCggA(T-NH2)GTTCTCgTG(2-aminopurine)TCCgATCT(ddC)-3'9603.463.42758009603.4632758000.0-0.40125'-(2'-F-G)GAACACCggA(Azide)GTTCTCgTG(2-aminopurine)ACCgATCT(FAM-dT)-3'10029.963.429730010029.9642973000.00.60135'-(2'-F-U)GAACACCggA(BiotinAzide)GTTCTCgTC(2-aminopurine)ACCgATCT(inverteddT)-3'9145.963.42775009145.962.82775000.0-0.60145'-(2,6-Diaminopurine)GAACACCggA(T-Biotin)GTTCTCgTC(2-aminopurine)GCCgATCT(TAMRA)-3'10185.165.630360010185.165.63036000.00.00155'-(NH2C6dT)GAACACCggA(T-FAM)GTTCTCgTT(2-aminopurine)CCCgATCA(2'-F-A)-3'10139.963.629760010139.963.12976000.0-0.50165'-(5-Br-dU)GAACACCggA(TAMRA)GTTCTCgTT(2-aminopurine)GCCgATCA(2'-F-C)-3'9249.8648653035000.00.90175'-(5-Me-dC)GAACACCggG(2'-F-A)CTTCTCgTA(2-aminopurine)AC9524.263.82859009524.2652859000.01.20

NO碱基序列FM-MWFM-TM(℃)FM-εIDT-MWIDT-TM(℃)IDT-εMW差值TM差值ε差值CgATCA(2'-F-G)-3'185'-(BiotindT)GAACACCggG(2'-F-C)CTTCTCgTG(2-aminopurine)GCCgATCA(2'-F-U)-3'9874.667.52748009874.668.52748000.01.00195'-(2'-F-A)CAACACCggG(2'-F-G)CTTCTCgTC(2-aminopurine)CCCgATCA(2,6-Diaminopurine)-3'9491.267.32724009491.269.52724000.02.20205'-(2'-F-C)CAACACCggG(2'-F-U)CTTCTCgTT(2-aminopurine)TCCgATCA(5-MedC)-3'9403.164.32663009403.164.32663000.00.00215'-(2'-F-G)CAACACCggG(2,6-Diaminopurine)CTTCTCgTA(Uni-LinkTMAminoMod)GCCgATCA(5-OctadiynyldU)-3'9305.166.02751009305.165.92751000.0-0.10225'-(2'-F-U)CAACACCggG(5-Br-dU)CTTCTCgTG(Uni-LinkTMAminoMod)CCCgATCA(3'NH2C6-dT)-3'8858.667.72647008858.666.22647000.0-1.50235'-(2,6-Diaminopurine)CAACACCggG(5-Me-dC)CTTCTCgTC(Uni-LinkTMAminoMod)TCCgATCA(BiotindT)-3'9502.565.82633009502.567.12633000.01.30245'-(NH2C6dT)CAACACCggG(5-OctadiynyldU)CTTCTCgTT(Uni-LinkTMAminoMod)ACCgATCA(DABCYL)-3'9495.564.02780839495.564.22780830.00.20255'-(5-Br-dU)CAACACCggG(5-TA8807.665.82970008807.665.52970000.0-0.30

NO碱基序列FM-MWFM-TM(℃)FM-εIDT-MWIDT-TM(℃)IDT-εMW差值TM差值ε差值MRATMAzide)CTTCTCgTA(Uni-LinkTMAminoMod)CCCgATCA(ddC)-3'265'-(5-Me-dC)CAACACCggG(6-FAMTMAzide)CTTCTCgTA(Uni-LinkTMAminoMod)TCCgATCA(FAM-dT)-3'9300.363.73012009300.364.13012000.00.40275'-(BiotindT)CAACACCggG(T-NH2)CTTCTCgTG(Uni-LinkTMAminoMod)TCCgATCA(inverteddT)-3'9643.666.12731009643.665.32731000.0-0.80285'-(2'-F-A)TAACACCggG(Azide)CTTCTCgTG(Uni-LinkTMAminoMod)ACCgATCA(TAMRA)-3'9436.564.83138009436.562.53138000.0-2.30295'-(2'-F-C)TAACACCggG(BiotinAzide)CTTCTCgTC(Uni-LinkTMAminoMod)ACCgATCG(2'-F-A)-3'8834.765.72761008834.763.42761000.0-2.30305'-(2'-F-G)TAACACCggG(T-Biotin)CTTCTCgTC(Uni-LinkTMAminoMod)GCCgATCG(2'-F-C)-3'9551.467.12717009551.466.72717000.0-0.40315'-(2'-F-U)TAACACCggG(T-FAM)CTTCTCgTT(Uni-LinkTMAminoMod)CCCgATCG(2'-F-G)-3'9659.465.82857009659.463.32857000.0-2.50325'-(2,6-Diaminopurine)TAACACCggG(TAMRA)CTTCTCgTT(Uni-LinkTMAminoMod)GCCgATCG(2'-F-U)-3'8893.866.22995008893.864.32995000.0-1.90335'-(NH2C69343.261.72790009343.262.12790000.00.40

NO碱基序列FM-MWFM-TM(℃)FM-εIDT-MWIDT-TM(℃)IDT-εMW差值TM差值ε差值dT)TAACACCggC(2'-F-A)TTTCTCgTA(Uni-LinkTMAminoMod)ACCgATCG(2,6-Diaminopurine)-3'345'-(5-Br-dU)TAACACCggC(2'-F-C)TTTCTCgTG(Uni-LinkTMAminoMod)GCCgATCG(5-MedC)-3'9206.866.22595009206.868.12595000.01.90355'-(5-Me-dC)TAACACCggC(2'-F-G)TTTCTCgTC(Uni-LinkTMAminoMod)CCCgATCG(5-OctadiynyldU)-3'9192.065.82647009192.066.32647000.00.50365'-(BiotindT)TAACACCggC(2'-F-U)TTTCTCgTT(Uni-LinkTMAminoMod)TCCgATCG(3'NH2C6-dT)-3'9170.262.12668009170.261.92668000.0-0.20375'-(FluoresceindT)AAACACCggC(2,6-Diaminopurine)TTTCTCgTA(spacer18)GCCgATCG(BiotindT)-3'10455.464.228880010455.464.32888000.00.10385'-(FluoresceindT)GAACACCggC(5-Br-dU)TTTCTCgTG(spacer18)CCCgATCG(DABCYL)-3'10235.966.527788310235.968.52778830.02.00395'-(FluoresceindT)CAACACCggC(5-Me-dC)TTTCTCgTC(spacer18)TCCgATCG(ddC)-3'9915.864.82724009915.867.72724000.02.90405'-(FluoresceindT)TAACACCggC(5-Octadiynyl10589.461.729710010589.462.82971000.01.10

NO碱基序列FM-MWFM-TM(℃)FM-εIDT-MWIDT-TM(℃)IDT-εMW差值TM差值ε差值dU)TTTCTCgTT(spacer18)ACCgATCG(FAM-dT)-3'415'-(6-FAM)AAACACCggC(5-TAMRATMAzide)TTTCTCgTA(spacer18)CCCgATCG(inverteddT)-3'9366.364364.73159600.00.60425'-(6-FAM)AAACACCggC(6-FAMTMAzide)TTTCTCgTA(spacer18)TCCgATCG(TAMRA)-3'9669.762.03323609669.763.13323600.01.10435'-(6-FAMTMAzide)AAACACCggC(T-NH2)TTTCTCgTG(spacer18)TCCgATCC(2'-F-A)-3'9305.264.32821009305.263.42821000.0-0.90445'-(AcryditeTM)AAACACCggC(Azide)TTTCTCgTG(spacer18)ACCgATCC(2'-F-C)-3'9124.264.32632009124.264.82632000.00.50455'-(AlexaFluor®488)AAACACCggC(BiotinAzide)TTTCTCgTC(spacer18)ACCgATCC(2'-F-G)-3'9527.464.33037009527.463.73037000.0-0.60465'-(AlexaFluor®532)AAACACCggC(T-Biotin)TTTCTCgTC(spacer18)GCCgATCC(2'-F-U)-3'10281.366.629950010281.365.32995000.0-1.30475'-(AlexaFluor®405)AAACACCggC(T-FAM)TTTCTCgTT(spacer18)CCCgATCC(2,6-Diaminopurine)-3'9650.664.42698009650.665.12698000.00.70485'-(AlexaFluor®594)AAACACCggC(TAMRA)TTTCTCgTT(spacer18)GCCgATCC(5-MedC)-3'9702.864.93285009702.8663285000.01.10

NO碱基序列FM-MWFM-TM(℃)FM-εIDT-MWIDT-TM(℃)IDT-εMW差值TM差值ε差值495'-(AlexaFluor®647)AAACACCggT(2'-F-A)ATTCTCgTA(spacer18)ACCgATCC(5-OctadiynyldU)-3'10623.260.331180010623.2613118000.00.70505'-(AlexaFluor®660)AAACACCggT(2'-F-C)ATTCTCgTG(spacer18)GCCgATCC(3'NH2C6-dT)-3'10036.964.130500010036.965.93050000.01.80515'-(NH2C6)AAACACCggT(2'-F-G)ATTCTCgTC(spacer18)CCCgATCC(BiotindT)-3'9516.563.72664009516.566.12664000.02.40525'-(NH2C12)AAACACCggT(2'-F-U)ATTCTCgTT(spacer18)TCCgATCC(DABCYL)-3'9548.660.62722839548.661.92722830.01.30535'-(NH2C6)AAACACCggT(2,6-Diaminopurine)ATTCTCgTA(Spacer9)GCCgATCC(ddC)-3'9048.061.62696009048.063.22696000.01.60545'-(Azide)AAACACCggT(5-Br-dU)ATTCTCgTG(Spacer9)CCCgATCC(FAM-dT)-3'9870.664.12739009870.664.62739000.00.50555'-(BIOTIN)AAACACCggT(5-Me-dC)ATTCTCgTC(Spacer9)TCCgATCC(inverteddT)-3'9380.361.42624009380.3642624000.02.60565'-(BiotinAzide)AAACACCggT(5-OctadiynyldU)ATTCTCgTT(Spacer9)ACCgATCC(TAMRA)-3'9378.460.32992009378.4612992000.00.70575'-(Biotin-TEG)AAACACCggT(5-9283.361.22993009283.361.12993000.0-0.10

NO碱基序列FM-MWFM-TM(℃)FM-εIDT-MWIDT-TM(℃)IDT-εMW差值TM差值ε差值TAMRATMAzide)ATTCTCgTA(Spacer9)CCCgATCA(2-Aminopurine)-3'585'-(C3Spacer)AAACACCggT(6-FAMTMAzide)ATTCTCgTA(Spacer9)TCCgATCA(5-Nitroindole)-3'8893.859859.43016000.00.30595'-(CY3)AAACACCggT(T-NH2)ATTCTCgTG(Spacer9)TCCgATCA(6-FAM)-3'10252.361.529380010252.361.42938000.0-0.10605'-(CY5.5)AAACACCggT(Azide)ATTCTCgTG(Spacer9)ACCgATCA(AlexaFluor®488)-3'10496.761.533390010496.761.43339000.0-0.10615'-(CY5)AAACACCggT(BiotinAzide)ATTCTCgTC(Spacer9)ACCgATCA(AlexaFluor®532)-3'10007.361.531800010007.361.43180000.0-0.10625'-(Dig)AAACACCggT(T-Biotin)ATTCTCgTC(Spacer9)GCCgATCA(AlexaFluor®532)-3'10611.963.131940010611.963.33194000.00.20635'-(dSpacer)AAACACCggT(T-FAM)ATTCTCgTT(Spacer9)CCCgATCA(AlexaFluor®594)-3'10272.261.632110010272.261.53211000.0-0.10645'-(DualBiotin)AAACACCggT(TAMRA)ATTCTCgTT(Spacer9)GCCgATCA(AlexaFluor®647)-3'11187.162.132720011187.1623272000.0-0.10

NO碱基序列FM-MWFM-TM(℃)FM-εIDT-MWIDT-TM(℃)IDT-εMW差值TM差值ε差值655'-(HEX)AAACACCggA(2'-F-A)CTTCTCgTA(Spacer9)ACCgATCA(AlexaFluor®660)-3'10649.961.434280010649.962.13428000.00.70665'-(IowaBlackFQ)AAACACCggA(2'-F-C)CTTCTCgTG(Spacer9)GCCgATCA(AlexaFluor®660)-3'9913.865.33137449913.867.53137440.02.20675'-(IowaBlackRQ)AAACACCggA(2'-F-G)CTTCTCgTC(Spacer9)CCCgATCA(NH2C6)-3'8708.764.93123578708.767.23123570.02.30685'-(JOE)AAACACCggA(2'-F-U)CTTCTCgTT(Spacer9)TCCgATCA(Azide)-3'9658.461.82830009658.461.52830000.0-0.30695'-(PCBiotin)AAACACCggA(2,6-Diaminopurine)CTTCTCgTA(dSpacer)GCCgATCA(BIOTIN)-3'9321.362.82657009321.3632657000.00.20705'-(PCSpacer)AAACACCggA(5-Br-dU)CTTCTCgTG(dSpacer)CCCgATCA(BHQ1)-3'9293.065.32647009293.064.82647000.0-0.50715'-(PO4)AAACACCggA(5-Me-dC)CTTCTCgTC(dSpacer)TCCgATCA(BHQ2)-3'9284.162.62665009284.164.22665000.01.60725'-(RhodamineGreen)AAACACCggA(5-OctadiynyldU)CTTCTCgTT(dSpacer)ACCgATCA(spacerC3)-3'9436.361.42846009436.361.12846000.0-0.30

NO碱基序列FM-MWFM-TM(℃)FM-εIDT-MWIDT-TM(℃)IDT-εMW差值TM差值ε差值735'-(RhodamineRed)AAACACCggA(5-TAMRATMAzide)CTTCTCgTA(dSpacer)CCCgATCA(Cholesterol)-3'9109.462.63280009109.462.53280000.0-0.10745'-(ROX)AAACACCggA(6-FAMTMAzide)CTTCTCgTA(dSpacer)TCCgATCA(Cy3)-3'9857.161.23108009857.160.83108000.0-0.40755'-(TAMRA)AAACACCggA(T-NH2)CTTCTCgTG(dSpacer)TCCgATCA(Cy5.5)-3'10502.762.931620010502.762.83162000.0-0.10765'-(TET)AAACACCggA(Azide)CTTCTCgTG(dSpacer)ACCgATCA(Cy5)-3'10180.062.929540010180.062.82954000.0-0.10775'-(TEX615)AAACACCggA(BiotinAzide)CTTCTCgTC(dSpacer)ACCgATCA(Digoxin)-3'8353.562.92888008353.562.82888000.0-0.10785'-(TexasRed-X)AAACACCggA(T-Biotin)CTTCTCgTC(dSpacer)GCCgATCA(DTPA)-3'9936.265.12779009936.264.72779000.0-0.40795'-(SHC6)AAACACCggA(T-FAM)CTTCTCgTT(dSpacer)CCCgATCA(dSpacer)-3'9537.463.02758009537.462.92758000.0-0.10805'-(Uni-LinkTMAminoModifier)AAACACCggA(TAMRA)CTTCTCgTT(dSpacer)GCCgATCA(dSpacer)-3'8564.663.52929008564.663.42929000.0-0.10815'-(6-FAM9927.559.83182609927.5603182600.00.20

NO碱基序列FM-MWFM-TM(℃)FM-εIDT-MWIDT-TM(℃)IDT-εMW差值TM差值ε差值ABI)AAACACCggA(2'-F-A)TTTCTCgTA(dSpacer)ACCgATCA(JOE)-3'825'-(DyLight800)AAACACCggA(2'-F-C)TTTCTCgTG(dSpacer)GCCgATCA(PO4)-3'9925.763.62654009925.765.72654000.02.10835'-(DyLight800)AAACACCggA(2'-F-G)TTTCTCgTC(dSpacer)CCCgATCA(RhodamineGreen)-3'10341.263.128330010341.264.72833000.01.60845'-(NH2C12)AAACACCggA(2'-F-U)TTTCTCgTT(dSpacer)TCCgATCA(RhodamineRed)-3'8945.960.12967008945.959.32967000.0-0.80855'-(NH2C3)AAACACCggA(2,6-Diaminopurine)TTTCTCgTA(SpacerC3)GCCgATCA(Spacer18)-3'9206.161.72676009206.161.62676000.0-0.10865'-(NH2C6)AAACACCggA(5-Br-dU)TTTCTCgTG(SpacerC3)CCCgATCA(Spacer9)-3'8923.663.62576008923.663.82576000.00.20875'-(5-FAM)AAACACCggA(5-Me-dC)TTTCTCgTC(SpacerC3)TCCgATCA(TEX)-3'9833.361.52910609833.362.82910600.01.30885'-(Alkyne)AAACACCggA(5-OctadiynyldU)TTTCTCgTT(SpacerC3)ACCgATCA(TexasRed-X)-3'9906.059.82819009906.0602819000.00.20895'-(AMCA)AAACACCggA(5-TAMRATMAzide)TTTCTCgTA(SpacerC3)CCCgATCA(SHC3)-3'8874.961.62969008874.961.52969000.0-0.10

NO碱基序列FM-MWFM-TM(℃)FM-εIDT-MWIDT-TM(℃)IDT-εMW差值TM差值ε差值905'-(AMCA)AAACACCggA(6-FAMTMAzide)TTTCTCgTA(SpacerC3)TCCgATCA(2,2'-Dipicolylamine)-3'9070.859.52842009070.859.82842000.00.30915'-(Bodipy493/503)AAACACCggA(T-NH2)TTTCTCgTG(SpacerC3)TCCgATCC(2-Aminopurine)-3'9586.362.12609009586.362.32609000.00.20925'-(DyLight750)AAACACCggA(Azide)TTTCTCgTG(SpacerC3)ACCgATCC(5-Nitroindole)-3'10074.162.129830010074.162.32983000.00.20935'-(Ce6)AAACACCggA(BiotinAzide)TTTCTCgTC(SpacerC3)ACCgATCC(6-FAM)-3'9630.762.12804009630.762.32804000.00.20945'-(Cholestrol)AAACACCggA(T-Biotin)TTTCTCgTC(SpacerC3)GCCgATCC(AlexaFluor®488)-3'10381.664.329880010381.664.12988000.0-0.20955'-(COOH)AAACACCggA(T-FAM)TTTCTCgTT(SpacerC3)CCCgATCC(AlexaFluor®532)-3'10148.262.231070010148.262.43107000.00.20965'-(Dylight680)AAACACCggA(TAMRA)TTTCTCgTT(SpacerC3)GCCgATCC(AlexaFluor®594)-3'10231.462.633020010231.462.83302000.00.20975'-(Dual5'BIOTIN)AAACACCggG(2'-F-A)TTTCTCgTA(SpacerC3)ACCgATCC(AlexaFluor®9557.662662.23117000.00.10

NO碱基序列FM-MWFM-TM(℃)FM-εIDT-MWIDT-TM(℃)IDT-εMW差值TM差值ε差值594)-3'985'-(Dylight680)AAACACCggG(2'-F-C)TTTCTCgTG(SpacerC3)GCCgATCC(AlexaFluor®647)-3'11060.766.628750011060.7682875000.01.40995'-(Dylight680)AAACACCggG(2'-F-G)TTTCTCgTC(SpacerC3)CCCgATCC(AlexaFluor®660)-3'10820.766.229220010820.767.22922000.01.001005'-(FAM)AAACACCggG(2'-F-U)TTTCTCgTT(SpacerC3)TCCgATCC(AlexaFluor®594)-3'10053.962.431956010053.961.43195600.0-1.001015'-(Ferrocene)AAACACCggG(2,6-Diaminopurine)TTTCTCgTAGCCgATCC(NH2C6)-3'8969.864.12591008969.863.62591000.0-0.501025'-(FITC)AAACACCggG(5-Br-dU)TTTCTCgTG)CCCgATCC(Azide)-3'9426.366.62694609426.3662694600.0-0.601035'-(Fluorescein)AAACACCggG(5-Me-dC)TTTCTCgTCTCCgATCC(BIOTIN)-3'9581.663.92704609581.665.32704600.01.401045'-(Hexynyl)AAACACCggG(5-OctadiynyldU)TTTCTCgTTACCgATCC(BHQ1)-3'9304.362.12664009304.362.32664000.00.201055'-(MB)AAACACCggG(5-TAMRATMAzide)TTTCTCgTACCCgATCC(B9326.563.92958009326.563.72958000.0-0.20

NO碱基序列FM-MWFM-TM(℃)FM-εIDT-MWIDT-TM(℃)IDT-εMW差值TM差值ε差值HQ2)-3'1065'-(N3)AAACACCggG(6-FAMTMAzide)TTTCTCgTATCCgATCC(spacerC3)-3'8623.761.82751008623.7622751000.00.201075'-(Nap)AAACACCggG(T-NH2)TTTCTCgTGTCCgATCC(Cholesterol)-3'10063.364.225740010063.3642574000.0-0.201085'-(NH2)AAACACCggG(Azide)TTTCTCgTGACCgATCC(Cy3)-3'9484.864.22658009484.8642658000.0-0.201095'-(Pyrene)AAACACCggG(BiotinAzide)TTTCTCgTCACCgATCC(Cy5.5)-3'9698.064.22784009698.0642784000.0-0.201105'-(rApp)AAACACCggG(T-Biotin)TTTCTCgTCGCCgATCC(Cy5)-3'10107.266.426530010107.265.82653000.0-0.601115'-(Rhamine123)AAACACCggG(T-FAM)TTTCTCgTTCCCgATCC(Digoxin)-3'9560.064.32806009560.064.12806000.0-0.201125'-(RhamineGreen)AAACACCggG(TAMRA)TTTCTCgTTGCCgATCC(DTPA)-3'8746.864.83027008746.864.63027000.0-0.201135'-(Rhamine)AAACACCggG(2'-F-A)ATTCTCgTAACCgATCC(dSpacer)-3'9420.461.82720009420.462.32720000.00.501145'-(Rhamine6G)AAACACCggG(2'-F-C)ATTCTCgTGGCCgATCC(HEX)-3'9917.466.42918009917.467.82918000.01.401155'-(SH)AAACACCggG(2'-F-G)ATTCTCgTCCCCgATCC(JOE)-3'9375.165.92796009375.167.52796000.01.601165'-(TexasRed)AAACACCggG(2'-F-U)ATTCTCgTTTCCgATCC(PO4)-3'9352.362.22751009352.361.72751000.0-0.50

NO碱基序列FM-MWFM-TM(℃)FM-εIDT-MWIDT-TM(℃)IDT-εMW差值TM差值ε差值1175'-(TRITC)AAACACCggG(2,6-Diaminopurine)ATTCTCgTA(Azobenzene)GCCgATCC(RhodamineGreen)-3'10120.163.830360010120.163.63036000.0-0.201185'-(XRITC)AAACACCggG(5-Br-dU)ATTCTCgTG(Azobenzene)CCCgATCC(RhodamineRed)-3'9676.666.43090009676.665.43090000.0-1.001195'-(