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实时荧光定量 PCR 引物设计原则
发布时间:2018-12-17   点击次数:263次

SHISHIYINGGUANGDINGLIANG PCR(real-time PCR)JISHUHEPUTONG PCR XIANGBI,ZHUNQUEXINGHELINGMINDUGAO,SUDUKUAI,XIAOLVGAO,JINNIANLAIBEIGUANGFANYINGYONGYUYIXUE、SHENGWUDENGGEGELINGYU,SHIJINXINGFENZISHENGWUXUEYANJIUBIBUKESHAODEJISHUGONGJU。ZAIKEYANFANGMIANKEDINGLIANGFENXIGEZHONGJIYINDEDEBIAODAFENXI,JIYINTUBIANHEDUOTAIXINGFENXI,DANHEGANSUANDUOTAI SNP CEDINGJIYIWEIJIYINDEJIANCE;ZAIYILIAOFANGMIANKEYONGYUMIANYIZUHUAFENXILINCHUANGJIBINGZAOQIZHENDUAN,BINGYUANTIJIANCE,NAIYAOXINGFENXI,ZHONGLIUWEIXIAOCANLIUBINGBIANYANJIU,XIBAOYINZIDEBIAODAFENXI,BINGDUGANRANDEDINGLIANGJIANCEDENG。

 

 

ZAIZHENGGEYINGGUANGDINGLIANG PCR SHIYANSHEJIZHONG,YINWUDESHEJISHISHIYANCHENGGONGDEGUANJIANYINSU,HENDACHENGDUJUEDINGLEJIANCEDELINGMINDUHEZHUNQUEXING,YIBAN real-time PCR YINWUDESHEJIYINGZUNXUNRUXIAYUANZE:

彩搜网1. YINWUYINGZAIHESUANXILIEBAOSHOUQUNEISHEJIBINGJUYOUTEYIXING。

彩搜网2. KUOZENGCHANWUZHANGDUZAI 80-150bp。ZHANGBUYAOCHAOGUO 300bp。

彩搜网3. CHANWUBUNENGXINGCHENGERJIJIEGOU(ZIYOUNENGXIAOYU 58.61KJ/mol)。

彩搜网4. YINWUZHANGDU:YIBANZAI 17-25 JIANJIZHIJIAN,SHANGXIAYOUYINWUBUYIXIANGCHATAIDA。YINWUZHANGDUSHIJUEDINGTUIHUOWENDU(Tm ZHI)ZHONGYAODEYINSU。YIBANLAIJIANG,MEIZENGJIAYIGEHEGANSUANKESHIYINWUTEYIXINGTIGAO 4 BEI,DANSHIYOUYUSHANGDEYUANYIN,YINWUYUEZHANG,TATUIHUOJIEHEDAOBA DNA SHANGXINGCHENGGONG DNA JUHEMEIJIEHEDEWENDINGSHUANGLIANMOBANDESULVYUEXIAO。

彩搜网5. YINWUZISHENBUNENGYOULIANXU 4 GEJIANJIDEHUBU,BIMIANXINGCHENGFAKAJIEGOU。

彩搜网6. YINWUZHIJIANBUNENGYOULIANXU 4 GEJIANJIDEHUBU,BIMIANXINGCHENGYINWUERJUTI。

7. YINWU G+C HANLIANGZAI 40%~60% ZHIJIAN,45-55% zuiHAO。YINWUZHONGJIANJIYAOSUIJIFENBU,JINLIANGJUNYUN,RUOSHIYINWUCUNZAIYANZHONGDE GC QINGXIANGHUO AT QINGXIANGZEKEYIZAIYINWU 5』DUANJIASHILIANGDE A、T HUO G、C WEIBALAIPINGHENGYIXIA。

8. YINWU Tm ZHIZAI 58-62 DUZHIJIAN,SHANGXIAYOUYINWU Tm ZHIBUYIXIANGCHATAIDA,zuiHAOBUYAOCHAOGUO 5 DU。RUGUOYINWUJIANJISHUJIAOSHAO,KEYISHIDANGTIGAOTUIHUOWENDU,TIGAO PCR DETEYIXING;RUGUOJIANJISHUJIAODUO,NEIMEKEYISHIDANGJIANDITUIHUOWENDU,SHI DNA SHUANGLIANJIEHE。

彩搜网9. YINWU 5′DUANKEYIXIUSHI;YINWU 5′DUANYIBANBUHUIYINGXIANGKUOZENGDETEYIXING,KEJINXINGXIUSHI,RUJIAMEIQIEWEIDIAN;BIAOJISHENGWUSU、YINGGUANG、DIGAOxin、Eu3+DENG;YINRUTUBIANWEIDIAN、CHARUYUQUESHITUBIANXULIEDENG。

10. YINWU 3′DUANBUKEXIUSHI,YINWU 3』DUANSHIYANSHENKAISHIDEDIFANG,JUEDINGKUOZENGDETEYIXING。3』DUANYINGBIKAILIANXUDE T/C,A/G(2-3 GE);YEBUNENGYOUXINGCHENGRENHEERJIJIEGOU。

彩搜网11. YINWU 3′DUANYAOBIKAIMIMAZIDEDI 3 WEI。YINWEIMIMAZIDEJIANBINGXING,YINMIMAZIDEDI 3 WEIYIFASHENGJIANBING,HUIYINGXIANGKUOZENGTEYIXINGYUXIAOLV。

 

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