醫(yī)療器械-軟件設(shè)計和開發(fā)-驗證報告-模板范例

醫(yī)療器械軟件設(shè)計開發(fā)驗證報告醫(yī)療器械軟件設(shè)計開發(fā)驗證報告PagePage#of70醫(yī)療器械軟件設(shè)計開發(fā)驗證報告記錄編號：目ODD錄DTOC\o"1-5"\h\z\o"CurrentDocument"\h第一部分總則 2\o"CurrentDocument"\h一、概述 2...\o"CurrentDocument"\h二、驗證方案 2.1、驗證目的 2.2、驗證范圍 2.\o"CurrentDocument"\h3、相關(guān)文件 2.4、職責(zé) 2.5、驗證條件 3.6、驗證方法 3.7、驗證合格標(biāo)準(zhǔn)及評價分析 7.8、驗證實施計劃 7.9、驗證周期 8.10、數(shù)據(jù)收集 8.\o"CurrentDocument"\h第二部分測試確認(rèn) 8\o"CurrentDocument"\h一、功能性測試： 8.\o"CurrentDocument"\h二、性能測試： 9.\o"CurrentDocument"\h三、可靠性和安全性測試 9\o"CurrentDocument"\h四、用戶場景測試 9.\o"CurrentDocument"\h五、準(zhǔn)確性測試 1. 0普通堿基測試 10\o"CurrentDocument"\h修飾堿基測試 16\o"CurrentDocument"\h兼并堿基測試 53\o"CurrentDocument"\h含RNA堿基測試 59含mRNA堿基測試 63\o"CurrentDocument"\h含特殊堿基I序列測試 65含特殊堿基U序列測試 67\o"CurrentDocument"\h第三部分結(jié)論 70第一部分總則一、概述本次驗證嚴(yán)格按照驗證方案要求實施，整個過程穩(wěn)定可控，驗證過程沒有出現(xiàn)偏差，F(xiàn)ilemaker的各項性能符合相關(guān)規(guī)定。二、驗證方案1、驗證目的通過驗證以證明使用Filemaker軟件設(shè)計的Filemaker數(shù)據(jù)庫系統(tǒng)在適用性、準(zhǔn)確性、穩(wěn)定性、可控性、安全性和保密性方面可以滿足公司質(zhì)量管理體系和用戶的要求與需要。2、驗證范圍使用Filemaker軟件設(shè)計的以下功能模塊：Filemaker.fp73、相關(guān)文件文件名稱文件描述DP_U_0025Filemaker操作方法QP_U_0114文件與資料的保密管理DP_U_0140Filemaker數(shù)據(jù)庫應(yīng)急方案與備份方案DP_U_0117Filemaker賬戶權(quán)限說明4、職責(zé)驗證委員會負(fù)責(zé)驗證方案的審批；負(fù)責(zé)驗證的協(xié)調(diào)工作，以保證本驗證方案規(guī)定項目的順利實施；負(fù)責(zé)驗證數(shù)據(jù)及結(jié)果的審核；負(fù)責(zé)驗證報告的審批。驗證委員會名單：姓名部門職位職責(zé)驗證小組負(fù)責(zé)驗證方案的起草、修改；負(fù)責(zé)組織本驗證方案的實施；負(fù)責(zé)驗證數(shù)據(jù)的統(tǒng)計、分析、審核，報驗證委員會審核；負(fù)責(zé)驗證報告的編寫，并報驗證委員會。驗證小組名單：5、驗證條件FilemakerServerAdvance9.2,數(shù)據(jù)庫托管服務(wù)器端；WindowsServer2003,數(shù)據(jù)庫托管服務(wù)器端操作系統(tǒng);FilemakerPro11.0,數(shù)據(jù)庫客戶端；WindowsXP,數(shù)據(jù)庫客戶端操作系統(tǒng)；InternetExplorer7.0,WebPublishing訪問客戶端。6、驗證方法質(zhì)量策劃風(fēng)險管理計劃見：《QR_U_0041_Filemaker軟件驗證報告》配制管理計劃見：《QR_U_0041_Filemaker軟件驗證報告》系統(tǒng)需求定義見《Filemaker數(shù)據(jù)庫需求分析》設(shè)計文檔見《Filemaker總體設(shè)計說明書》測試方法

功能性測試對FilemakerFilemaker數(shù)據(jù)庫的各項功能進(jìn)行測試，以驗證其適合性、互操作性和保密與安全性符合QMS的要求：特性測試內(nèi)容說明符合要求判定適合性文件的共享與托管，即時網(wǎng)絡(luò)發(fā)布使用FSA托管并發(fā)布數(shù)據(jù)庫，使用FilemakerPro和IE兩種方式進(jìn)行訪問查找所需數(shù)據(jù)分別測試一般搜索、限制搜索和擴(kuò)展搜索功能，導(dǎo)出搜索結(jié)果數(shù)據(jù)的導(dǎo)入導(dǎo)出批量導(dǎo)入，導(dǎo)出文件批量導(dǎo)入，導(dǎo)出文件信息數(shù)據(jù)庫布局的創(chuàng)建與設(shè)計根據(jù)需要更改布局、字段、腳本保密與安全性賬戶和權(quán)限集管理員可以控制增加任意數(shù)量的賬戶。測試系統(tǒng)及其數(shù)據(jù)訪問的可控制性。測試系統(tǒng)防止非法操作的模式，包括防止非授權(quán)的創(chuàng)建、刪除和修改程序或信息可控的記錄審核流程可增加或修改審核流程，可查看審核狀態(tài)自動備份可設(shè)置自動備份周期，自動備份數(shù)據(jù)庫限制編輯測試系統(tǒng)防止數(shù)據(jù)被誤刪、誤改和被破壞的能力測試者：年月日驗證小組確認(rèn)人簽字：年月日性能測試對Filemaker構(gòu)建的Filemaker數(shù)據(jù)庫系統(tǒng)的性能進(jìn)行強(qiáng)度測試，以驗證其系統(tǒng)響應(yīng)時間、系統(tǒng)存儲量和異常狀態(tài)系統(tǒng)性能符合QMS的要求：測試系統(tǒng)的響應(yīng)時間，包括單個用戶、多用戶并發(fā)的情況，批量導(dǎo)入記錄的響應(yīng)時間；測試系統(tǒng)的最大存儲量、100條記錄的平均存儲量；測試運行條件在異常狀態(tài)下，或在人為設(shè)定的狀態(tài)下，系統(tǒng)的性能。

特性測試內(nèi)容響應(yīng)時間系統(tǒng)響應(yīng)時間單客戶端訪問數(shù)據(jù)庫單IE瀏覽器訪問數(shù)據(jù)庫5個客戶端同時訪問數(shù)據(jù)庫10個IE瀏覽器同時訪問數(shù)據(jù)庫批量導(dǎo)入100條記錄的響應(yīng)時間系統(tǒng)存儲量數(shù)據(jù)庫最大存儲量100條記錄平均存儲量異常狀態(tài)系統(tǒng)性能服務(wù)器重啟后系統(tǒng)性能系統(tǒng)恢復(fù)后的性能域環(huán)境殺毒軟件掃描階段系統(tǒng)的性能測試者： 驗證小組確認(rèn)人簽字：年月日 年月日可靠性和安全性測試見：《QR_U_0041_Filemaker軟件驗證報告》用戶場景測試測試Filemaker.fp7的實際應(yīng)用場景，以用戶角色權(quán)限完成一項特定的業(yè)務(wù)處理流程測試其易理解性、易學(xué)性、易操作性和時間特性：特性測試內(nèi)容說明符合要求判定易理解性系統(tǒng)的各項功能是否容易被識別或被理解參考Filemakerintroduction-databasesolutionQR_U_0001_數(shù)據(jù)庫培訓(xùn)20100902界面的輸入和輸出的格式和含義是否容易被理解易學(xué)性系統(tǒng)的在線幫助是否容易定位，是否有效各數(shù)據(jù)庫負(fù)責(zé)人和管理員是否明確及聯(lián)絡(luò)方式是否有效測試用戶文檔的有效性測試“3、相關(guān)文件”中所有文件的有效性易操作性系統(tǒng)是否對輸入數(shù)據(jù)進(jìn)行有效性檢查以正確操作、誤操作模式、非常規(guī)操作

中斷執(zhí)行功能是否可以在動作完成之前被取消模式為框架設(shè)計測試用例。誤操作模式有錯誤的數(shù)據(jù)類型作參數(shù)、錯誤的輸入數(shù)據(jù)序列、錯誤的操作序列。確認(rèn)參數(shù)是否易于選擇，是否有缺省值解釋的消息是否明確界面提示是否有效確認(rèn)系統(tǒng)能否提示出錯的風(fēng)險、能否糾正錯誤的輸入、能否從錯誤中恢復(fù)確認(rèn)定制能力的有效性時間特性系統(tǒng)響應(yīng)時間完成一項規(guī)定的任務(wù)所需的時間平均響應(yīng)時間執(zhí)行若干并行任務(wù)所需的平均時間響應(yīng)極限時間最大負(fù)載條件下，系統(tǒng)完成某項任務(wù)需要時間的極限系統(tǒng)的吞吐量給定的時間周期內(nèi)系統(tǒng)能完成的任務(wù)數(shù)量平均吞吐量在一個單位時間內(nèi)系統(tǒng)處理并發(fā)任務(wù)的平均數(shù)極限吞吐量最大負(fù)載條件下，在給定的時間周期內(nèi)系統(tǒng)處理的最多并發(fā)任務(wù)數(shù)測試者：年月日驗證小組確認(rèn)人簽字：年月日準(zhǔn)確性測試隨機(jī)抽取100條普通堿基序列，驗證普通堿基的分子量、TM值和消光系數(shù)計算與IDT網(wǎng)站的差異率。所有兼并堿基對隨機(jī)組合，每條序列最多三對，不包含兼并堿基對位于序列首位的情況，驗證兼并堿基的分子量、TM值和消光系數(shù)計算與IDT網(wǎng)站的差異率。所有修飾堿基隨機(jī)組合，每條序列最多四種，序列中不包含兼并堿基、特殊堿基和IDT網(wǎng)站上沒有的修飾堿基，驗證修飾堿基的分子量、TM值和消光系數(shù)計算與IDT網(wǎng)站的差異率。隨機(jī)選取以下四種含RNA的序列個10條，驗證含RNA的序列的分子量、TM值和消光系數(shù)計算與IDT網(wǎng)站的差異率。全部為RNA5′端部分含RNA中間部分含RNA3′端部分含RNA隨機(jī)選取以下三種含mRNA的序列各5條，驗證含mRNA的序列的分子量、TM值和消光系數(shù)計算與IDT網(wǎng)站的差異率。5′端部分含RNA中間部分含RNA3′端部分含RNA6.5.5.5隨機(jī)選取以下三種含特殊堿基I的序列各5條，驗證含I的序列的TM值和消光系數(shù)計算與IDT網(wǎng)站的差異率。5′端部分含I中間部分含I3′端部分含I6.5.5.5隨機(jī)選取以下三種含特殊堿基U的序列各5條，驗證含U的序列的TM值和消光系數(shù)計算與IDT網(wǎng)站的差異率。5′端部分含U中間部分含U3′端部分含U7、驗證合格標(biāo)準(zhǔn)及評價分析Filemaker.fp7數(shù)據(jù)庫的適合性、互操作性和保密與安全性符合QMS的要求；Filemaker.fp7數(shù)據(jù)庫的系統(tǒng)響應(yīng)時間、系統(tǒng)存儲量和異常狀態(tài)系統(tǒng)性能應(yīng)符合用戶的要求；Filemaker.fp7數(shù)據(jù)庫的可靠性和安全性應(yīng)符合QMS的要求；Filemaker.fp7數(shù)據(jù)庫的易理解性、易學(xué)性、易操作性和時間特性應(yīng)能夠適應(yīng)用戶的要求；Filemaker.fp7數(shù)據(jù)庫的應(yīng)急方案應(yīng)能保證其安全性和可持續(xù)性。Filemaker.fp7數(shù)據(jù)庫的準(zhǔn)確性應(yīng)符合用戶要求。8、驗證實施計劃驗證步驟時間安排Filemaker.fp7導(dǎo)入2019-9-6準(zhǔn)確性驗證Filemaker.fp7試用2019-9-13到2019-9-31

9、驗證周期10、數(shù)據(jù)收集第二部分測試確認(rèn)、功能性測試：特性測試內(nèi)容說明符合要求判定適合性文件的共享與托管，即時網(wǎng)絡(luò)發(fā)布使用FSA托管并發(fā)布數(shù)據(jù)庫，使用FilemakerPro和IE兩種方式進(jìn)行訪問查找所需數(shù)據(jù)分別測試一般搜索、限制搜索和擴(kuò)展搜索功能，導(dǎo)出搜索結(jié)果數(shù)據(jù)的導(dǎo)入導(dǎo)出批量導(dǎo)入，導(dǎo)出文件批量導(dǎo)入，導(dǎo)出文件信息數(shù)據(jù)庫布局的創(chuàng)建與設(shè)計根據(jù)需要更改布局、字段、腳本保密與安全性賬戶和權(quán)限集管理員可以控制增加任意數(shù)量的賬戶。測試系統(tǒng)及其數(shù)據(jù)訪問的可控制性。測試系統(tǒng)防止非法操作的模式，包括防止非授權(quán)的創(chuàng)建、刪除和修改程序或信息可控的記錄審核流程可增加或修改審核流程，可查看審核狀態(tài)自動備份可設(shè)置自動備份周期，自動備份數(shù)據(jù)庫限制編輯測試系統(tǒng)防止數(shù)據(jù)被誤刪、誤改和被破壞的能力測試者：年月日驗證小組確認(rèn)人簽字：年月日

二、性能測試：特性測試內(nèi)容響應(yīng)時間系統(tǒng)響應(yīng)時間單客戶端訪問數(shù)據(jù)庫單IE瀏覽器訪問數(shù)據(jù)庫5個客戶端同時訪問數(shù)據(jù)庫10個IE瀏覽器同時訪問數(shù)據(jù)庫批量導(dǎo)入100條記錄的響應(yīng)時間系統(tǒng)存儲量數(shù)據(jù)庫最大存儲量100條記錄平均存儲量異常狀態(tài)系統(tǒng)性能服務(wù)器重啟后系統(tǒng)性能系統(tǒng)恢復(fù)后的性能域環(huán)境殺毒軟件掃描階段系統(tǒng)的性能測試者： 驗證小組確認(rèn)人簽字：年月日 年月日三、可靠性和安全性測試見：《QR_U_0041_Filemaker軟件驗證報告》四、用戶場景測試特性測試內(nèi)容說明符合要求判定易理解性系統(tǒng)的各項功能是否容易被識別或被理解參考Filemakerintroduction-databasesolutionQR_U_0001_數(shù)據(jù)庫培訓(xùn)20100902界面的輸入和輸出的格式和含義是否容易被理解易學(xué)性系統(tǒng)的在線幫助是否容易定位，是否有效各數(shù)據(jù)庫負(fù)責(zé)人和管理員是否明確及聯(lián)絡(luò)方式是否有效測試用戶文檔的有效性測試“3、相關(guān)文件”中所有文件的有效性易操作性系統(tǒng)是否對輸入數(shù)據(jù)進(jìn)行有效性檢查以正確操作、誤操作模式、非常規(guī)操作模式為框架設(shè)計測試用例。誤操作模式有錯誤的數(shù)據(jù)類型作參數(shù)、錯誤的輸入中斷執(zhí)行功能是否可以在動作完成之前被取消

特性測試內(nèi)容說明符合要求判定確認(rèn)參數(shù)是否易于選擇，是否有缺省值數(shù)據(jù)序列、錯誤的操作序列。解釋的消息是否明確界面提示是否有效確認(rèn)系統(tǒng)能否提示出錯的風(fēng)險、能否糾正錯誤的輸入、能否從錯誤中恢復(fù)確認(rèn)定制能力的有效性時間特性系統(tǒng)響應(yīng)時間完成一項規(guī)定的任務(wù)所需的時間平均響應(yīng)時間執(zhí)行若干并行任務(wù)所需的平均時間響應(yīng)極限時間最大負(fù)載條件下，系統(tǒng)完成某項任務(wù)需要時間的極限系統(tǒng)的吞吐量給定的時間周期內(nèi)系統(tǒng)能完成的任務(wù)數(shù)量平均吞吐量在一個單位時間內(nèi)系統(tǒng)處理并發(fā)任務(wù)的平均數(shù)極限吞吐量最大負(fù)載條件下，在給定的時間周期內(nèi)系統(tǒng)處理的最多并發(fā)任務(wù)數(shù)測試者：驗證小組確認(rèn)人簽字：年月日年月日五、準(zhǔn)確性測試1普通堿基測試1.1隨機(jī)抽取100條普通堿基序列，驗證普通堿基的分子量、TM值和消光系數(shù)計算與IDT網(wǎng)站的差異率。NO堿基序列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堿基序列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堿基序列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堿基序列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堿基序列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驗證者：年月日驗證小組確認(rèn)人簽字：年月日普通堿基分子量、Tm值和摩爾消光系數(shù)計算數(shù)據(jù)分析：通過與IDT網(wǎng)站計算數(shù)據(jù)的差值分析FM數(shù)據(jù)庫計算的準(zhǔn)確性：

差值分子量Tm值摩爾消光系數(shù)最大值00.40最小值0-0.40平均值00.10標(biāo)準(zhǔn)偏差00.1946840樣本數(shù)100100100設(shè)定量000下限規(guī)格0-0.50上限規(guī)格00.60做TM值差值的I-MR控制圖：UCL=0.556X=0.0521 11 2131UCL=0.556X=0.0521 11 213171 81 9141 51 61觀測值718191111213141 51 718191111213141 51 61觀測值結(jié)論：AFM數(shù)據(jù)庫普通堿基序列的分子量計算與IDT網(wǎng)站平均差值為0。AFM數(shù)據(jù)庫普通堿基序列的TM值計算與IDT網(wǎng)站平均差值為0.1℃，上限規(guī)格為0.6℃，下限規(guī)格為-0.5℃。AFM數(shù)據(jù)庫普通堿基序列的摩爾消光系數(shù)計算與IDT網(wǎng)站平均差值為0。

2修飾堿基測試所有修飾堿基隨機(jī)組合，每條序列最多四種，序列中不包含兼并堿基、特殊堿基和IDT網(wǎng)站上沒有的修飾堿基，驗證修飾堿基的分子量、TM值和消光系數(shù)計算與IDT網(wǎng)站的差異率。NO堿基序列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堿基序列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堿基序列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堿基序列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堿基序列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堿基序列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堿基序列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堿基序列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堿基序列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堿基序列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堿基序列FM-MWFM-TM(℃)FM-εIDT-MWIDT-TM(℃)IDT-εMW差值TM差值ε差值A(chǔ)BI)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堿基序列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堿基序列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堿基序列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堿基序列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'-(