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荧光-PCR法

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禽流感病毒H9N2亚型(AIV-H9N2)双重核酸检测试剂盒(荧光PCR法)

 

火马电竞  CHANPINMINGCHENG

 GUIGE

火马电竞  FENLEI

 禽流感病毒H9N2亚型(AIV-H9N2)双重核酸检测试剂盒(荧光PCR法)

 48T/HE

火马电竞  YINGGUANGJIANCESHIJIHE


 禽流感病毒H9N2亚型(AIV-H9N2)双重核酸检测试剂盒(荧光PCR法)火马电竞 原理是由一对引物介导,能在动植物体外对特定基因(DNA)片断进行快速酶促扩增,经过n个热循环扩增,扩增产物中所含特定基因数是原始模板数的(1+E)n(0<E<1,扩增效率=倍,使得检测扩增产物中的特定基因成为可能。

TEYIXINGQIANG

火马电竞 PCRFANYINGDETEYIXINGJUEDINGYINSUWEI:

①YINWUYUMOBANDNATEYIZHENGQUEDEJIEHE;

②JIANJIPEIDUIYUANZE;

③Taq DNAJUHEMEIHECHENGFANYINGDEZHONGSHIXING;

火马电竞       ④BAJIYINDETEYIXINGYUBAOSHOUXING。

CHANPINJITEDIAN :

火马电竞 JUHEMEILIANSHIFANYING(PCR)SHITIWAIMEICUHECHENGTEYIDNAPIANDUANDEYIZHONGFANGFA,YOUGAOWENBIANXING、DIWENTUIHUO(FUXING)JISHIWENYANSHENDENGJIBUFANYINGZUCHENGYIGEZHOUQI,XUNHUANJINXING,SHIMUDEDNADEYIXUNSUKUOZENG,JUYOUTEYIXINGQIANG、LINGMINDUGAO、CAOZUOJIANBIAN、SHENGSHIDENGTEDIAN。BENCHANPINJIUSHIWEIMANZUZHEIYIXUQIUGENJU PCR YUANLIKAIFADECHANPIN,TAJUYOUXIALIETEDIAN:

1. YIGUANSHICAOZUO,YONGHUZHIXUYAOTIGONGYANGPINJIKE。

火马电竞 2. GENJUDADOUREXIUKEDANBAIJIYINBAOSHOUQUYUSHEJIYINWU,NENGZHUANYIXINGJIANCECHUYANGPINZHONGDEDADOUCHENGFEN,DANBUNENGJIANCEQITAFEIDADOUCHENGFEN。

3. KUAISU,ZHENGGEJIANCEGUOCHENG(ANYIGEYANGPINJI)SUOXUSHIJIANJINWEI 2.0 XIAOSHI。

4. ZHIXUYAOPUTONG PCR YIHENINGJIAODIANYONGYI,WUXUPEIZHIGUIZHONGYIQISHEBEI。

5. DUIHUNHEYANGPINZHONGDADOUCHENGFENDEJIANCEXIAXIANWEI 0.1%,DUIYANGPINZHONGDADOUCHENGFENDEHESUANJIANCEXIAXIANWEI 1.0ng/μL。

6. BENZHINENGYONGYUKEYAN,ZUGOU 50 CI 40uL TIXIDECHANGGUI PCR。

 禽流感病毒H9N2亚型(AIV-H9N2)双重核酸检测试剂盒(荧光PCR法)具有下列特点:

火马电竞 1.GENJU ZHIBIAODEBAOSHOUXULIESHEJIDEZHUANYIXINGYINWU,YUXIANGGUANBINGDUWUJIAOCHAFANYING。

火马电竞 2.LINGMINDUBICHANGGUI PCR GAO 2-3 GESHULIANGJI,KEYIDADAOJIBAIKAOBEI/FANYING。

火马电竞 3.JIKAIJIYONG,YONGHUZHIXUYAOTIGONGYANGPINMOBAN,CAOZUOJIANDAN,DINGLIANGZHUNQUEKUAISU。

4.YIGUANSHIYINGGUANGDINGLIANG PCR JIANCE,BIMIANHOUXUWURAN。

5.BENSHIJIHEZUGOU 50 CI 20μL FANYINGTIXIDEYINGGUANGDINGLIANG PCR。

6.BENCHANPINZHISHIYONGYUKEYAN,BUNENGYONGYULINCHUANGZHENDUAN。

火马电竞 GUIGEJICHENGFEN                            CHENGFEN

2×qPCR MagicMix                 500 μL(ZHUANG A DAI)

WEILIANGHESUANXISHIYE(YINGGUANG PCR ZHUANYONG)1 mL(ZHUANG A DAI)

火马电竞 PCR YINWUHUNHEWU                        100 μL(ZHUANG A DAI)

火马电竞 JIYINYANGXINGDUIZHAO                           50 μL(ZHUANG B DAI)

DNA BINGDULIEJIEYE(SHIYONGZHUANG)       15 CI(9 mL)

SHIYONGSHOUCE                              1 FEN

火马电竞 YUNSHUJIBAOCUN:DIWENYUNSHU、-20℃BAOCUN(A DAISHIJIFANGYANGPINZHUNBEIQU、B DAIDEYANGXINGDUIZHAOFENKAIFANGZHI),

ZIBEISHIJIDNA MOBAN、10×ROX(GENJUJIXINGJUEDING,JUTIJIANSHIYONGFANGFA)。

YINGYONGANLI

 禽流感病毒H9N2亚型(AIV-H9N2)双重核酸检测试剂盒(荧光PCR法)样品 DNA 的制备

1.YONGZIXUANFANGFACHUNHUAYANGPINDE DNA,BENSHIJIHEGENSHICHANGSHANGDADUOSHUBINGDU DNA TIQUSHIJIHEJIAN RONG。。YEKEYIXUANGOUBENGONGSIDEYIGUANSHIBINGDU DNAout HUOQISHENGJIBANZHUSHIBINGDU DNAout。 BENSHIJIHEMIANFEIZENGSONG 15 CI DNA BINGDULIEJIEYESHIYONGZHUANG(YIGUANSHIBINGDU DNAout DECHENGFEN)。

XISHIYANGXINGDUIZHAO(YI 10E2-10E7 ZHEI 6 GE 10 BEIXISHIDUWEILI。YOUYUBIAOZHUNPINNONGDUFEICHANGGAO,YINCIXIALIEXISHICAOZUOYIDINGYAOZAIDULIDEQUYUJINXING,QIANWANBUNENGWURANYANGPINHUOBENSHIJIHEDEQITACHENGFEN)。WEIZENGJIACHANPINWENDINGXINGHEBIMIANKUOSANCHUANRANXINGBINGYUAN,BENCHANPINBUTIGONGHUOTIYANGPINZUOYANGXINGDUIZHAO,ZHITIGONGKEYIZHIJIESHIYONGDEZHANGWEI 86bp DE DNA PIANDUANZUOWEIYANGXINGDUIZHAO。

2.BIAOJI 6 GELIXINGUAN,FENBIEWEI 7,6,5,4,3,2。

3.YONGDAIXINQIANGTOUFENBIEJIARU 45 μL MOBANXISHIYE(YONGDAIXINQIANGTOU,XIATONG)。

4.ZAI 7 HAOGUANZHONGJIARU 5 μL 1×10E8 KAOBEI/μL DEYANGXINGDUIZHAO,CHONGFENZHENDANG 1 FENZHONG,DE1×10E7 KAOBEI/μL DEYANGXINGDUIZHAO。FANGBINGSHANGDAIYONG。5.HUANQIANGTOU,ZAI 6 HAOGUANZHONGJIARU 5 μL 1×10E7 KAOBEI/μL DEYANGXINGDUIZHAO(SHANGBUXISHISUODE),CHONGFENZHENDANG 1 FENZHONG,DE 1×10E6 KAOBEI/μL DEYANGXINGDUIZHAO。FANGBINGSHANGDAIYONG。

火马电竞 6.HUANQIANGTOU,ZAI 5 HAOGUANZHONGJIARU 5 μL 1×10E6 KAOBEI/μL DEYANGXINGDUIZHAODAO 5 HAOGUANZHONG,CHONGFENZHENDANG 1 FENZHONG,DE 1×10E5 KAOBEI/μL DEYANGXINGDUIZHAO。FANGBINGSHANGDAIYONG。

火马电竞 7.HUANQIANGTOU,ZAI 4 HAOGUANZHONGJIA 5 μL 1×10E5 KAOBEI/μL DEYANGXINGDUIZHAODAO 4 HAOGUANZHONG,DE1×10E4 KAOBEI/μL DEYANGXINGDUIZHAO。FANGBINGSHANGDAIYONG。8.ZHONGFUSHANGMIANDECAOZUOZHIDAODEDAO 6 GEXISHIDUDEYANGXINGDUIZHAO。FANGBINGSHANGDAIYONG。

火马电竞 BAOPAOZHENYANGBINGDUPCRSHIJIHESHEZHI qPCR FANYING(20  μL TIXI,ZAIYANGPINZHIBEISHIJINXING)

火马电竞 9.ZAI PCR GUANZHONGJIARUXIALIECHENGFEN(DAICEYANGPINXUYAOZUOSANCIZHONGFU,XIABIAOZHILIECHUYICI。YANGPINGUANHEYINXINGDUIZHAOSHEZHIWANBIHOUCAISHEZHIYANGXINGDUIZHAO,BINGQIEYANGXINGDUIZHAOYANGPINYAODENGSUOYOUGUANZIGAISHANGGAIZICHUCUNHAOHOU*HOUJIA):

ZHU:JIN ABI7500、7700 HE 7900 YIQIXUYAOSHIYONG ROX ZUOWEIDUIZHAO,QITAYINGGUANG PCR YIQI(RU iCycler IQ、MJ Option、MJ Chromo4、MX3000、MX4000、RotorGene3000、RotorGene 6000 HE LightCycler480)BUXUYAOSHIYONG ROX,ZEYONGSHUITIDAI。

10. GAISHANGGAIZIHOUSHANGJI,ANXIAMIANCANSHUJINXING PCR(JUTI PCR CANSHUKEYIGENJUYIQIBUTONGERZIXING

YOUHUA)。

GUOCHENGWENDUSHIJIAN

YUBIANXING95℃1 FENZHONG

PCR FANYING95℃15 MIAO

火马电竞 (30 GEXUNHUAN)60℃15 MIAO

72℃15 MIAO

11. SHUJUCAIJI

火马电竞 JUTICAOZUOANSUOYONGYIQITUIJIANDELIUCHENGJINXING。BENCHANPINZHONGSUOHANDEYINGGUANGRANLIAOZAIBUJIEHE DNA SHI,

XISHOUGUANGPUZAI 471 nm,JIEHE DNA SHIDEXISHOUGUANGPUZAI 500 nm,FASHEGUANG

PUZAI 530 nm。

SI、SHUJUCHULI

12. YIYANGXINGDUIZHAONONGDUDE log ZHIWEIHENGZHOU,YI Ct ZHIWEIZONGZHOU,HUIZHIBIAOZHUNQUXIAN。ZAIYIDAICEYANGPIN

火马电竞 DE Ct ZHICONGBIAOZHUNQUXIANSHANGTUISUANCHUYANGPIN DNA NONGDUDE log ZHI,ZAITUISUANCHUQINONGDU。


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