如何选择合适的腺相关病毒文件下载

 

病毒如何选择-腺病毒or慢病毒?

病毒的转导效率比常规真核表达载体高很多,因此特别适合于介导外源基因在难转染甚至无法转染的哺乳动物细胞中表达。现在常用的病毒有慢病毒、腺病毒、逆转录病毒和腺相关病毒,我们目前接的项目主要是做慢病毒和腺病毒,这两种病毒在提高基因表达量,缩短实验周期上各有优势,但是有时可能因为实验目的不同,需要选择合适的腺相关病毒进行实验研究,具体选择何种病毒要可根据这两种病毒各自的特点来决定:

MANBINGDU:

YOUDIAN:

KEYICHARUXIBAOJIYINZU,WENDINGBIAODA,CHIXUSHIJIANZHANG;  

BAOZHUANGZHOUQIDUAN(YIBANYAOLIANGGEYUE);

QUEDIAN:

GANRANXIAOLVXIANGDUIYUXIANBINGDUYAODIHENDUO;

BUNENGKUOZENG,YICIBAOZHUANGDEBINGDUYONGWANHOURUGUOZAIXUYAOZHINENGZHONGXINBAOZHUANG,CHENGBENGAO。

YICIBAOZHUANGSUODEDAODEDEBINGDUDIDUYEXIANGDUIJIAODI(10DE8CIFANGpfu/ml),BUDASHIHEZHIJIEYONGYUDONGWUSHIYAN;

XIANBINGDU:

YOUDIAN:

GANRANXIAOLVGAO,HENDUOXIBAODOUNENGDADAOJIN99%;

CHUNHUAHOUDEXIANBINGDUKEYIZHIJIEJINXINGDONGWUHUOTIZHUSHE;

KEYIZAITIWAIKUOZENG,MEICISHIYONGWANHOU,KEYIZIJIYONGBAOZHUANGXIBAOJINXINGKUOZENG,JIEYUECHENGBEN;

YICIBAOZHUANGSUODEDAODEDIDUJIAOGAO(CHUNHUAHOUWEI10DE11CIFANGpfu/ml,WEICHUNHUADEWEI10DE10CIFANGpfu/ml);

QUEDIAN: 

BUZHENGHEDAOJIYINZU,WUFAWENDINGBIAODA;

BIAODASHIJIANXIANGDUIYUMANBINGDUJIAODUAN(LIANGSANZHOUZHEIYANG)。BAOZHUANGXIANBINGDUDEZHOUQIHUIBIMANBINGDUZHANGYIXIE(YIBANYAOLIANGGEBANYUE),YINWEIXIANBINGDUBAOZHUANGGUOCHENGXIANGDUIJIAOFANSUO。

DANSHIZONGTILAISHUO,DUIYUHENDUOSHIYANLAISHUO,XUANZEXIANBINGDUYOUSHIGENGDA,BINGQIEHENDUOQINGKUANGXIAXIANBINGDUSHIKEYITIDAIMANBINGDULAIJINXINGSHIYANDE:

YI、HOUXUSHIYANZHOUQIBUTAIZHANG(YIGEYUENEI)。XUANZEXIANBINGDUBIMANBINGDUXIAOLVGENGGAO,YONGMANBINGDUDEHUAGANRANXIAOLVBUHUITAIGAO,XIAOLVGAODEHUAYEJIU70%DAO80%ZHEIYANG,BUNENGZHIJIEJINXINGHOUXUSHIYAN,XUYAOSHAIXUANWENDINGXIBAOZHU,ERSHAIXUANWENDINGXIBAOZHUSHIJIANZHISHAOXUYAOLIANGZHOUSHIJIAN,SHAIXUANGUOCHENGBUSHUNLIDEHUA,XUYAODESHIJIANGENGDUO,HUIDANWUSHIYAN。ERXIANBINGDUGANRANXIAOLVGAO,HENDUOXIBAODEXIAOLVKEYIDADAO99%YISHANG,ZHEIYANGJIUBUXUYAOHUAFEIEWAIDESHIJIANSHAIXUANWENDINGXIBAOZHU,KEZHIJIEJINXINGHOUXUSHIYAN,DUIYUSHIYANSHIJIANHEJINGLISHANGJIESHENGHENDUO。

ER、MUDEXIBAOGANRANNANDUDA,QIEHOUXUSHIYANBUXUYAOJIANLIWENDINGXIBAOZHU。DUIYUZHEIZHONGQINGKUANG,DANGRANSHIQIANGLIEJIANYIYONGXIANBINGDU,BUSHUOXIANBINGDUGANRANXIAOLVHUIGAOHENDUO,CHENGBENSHANGYEHUIJIESHENGHENDUO。BIRUSHENJINGXIBAO,MANBINGDUGANRANXIAOLVGUJIZHIYOU30%ZHEIYANG,YAOBAGANRANSHANGDEXIBAOFENXUANCHULAIJIXUPEIYANGCAINENGJINXINGHOUXUSHIYAN,DANSHIHENDUOSHENJINGXIBAO(ZHONGLIUXIBAOCHUWAI)DOUSHIBUKECHUANDAIDE,RUGUOSHAIXUANHOUJIHUSHENGBULEDUOSHAOXIBAO,KENENGJIUWUFAWANCHENGHOUXUSHIYAN。DANSHIRUGUOYONGXIANBINGDUJIUBUHUICHUXIANZHEIYANGDEQINGKUANG,XIANBINGDUGANRANXIAOLVHUIBIMANBINGDUGAOHENDUO(KEYIDADAO70%ZUOYOU),ZHEIYANGSHAIXUANCHULAIDEXIBAOKEYIZHIJIEJINXINGHOUXUSHIYAN,JIESHENGLESHIJIANCHENGBEN。

SAN、YAOJINXINGDONGWUSHIYAN。DONGWUSHIYANXUYAODEBINGDULIANGDA,QIEBINGDUDEDIDUYAOHENGAO,ZHEIYANGCAINENGDADAOZUIXIAOTIJIZHUSHEZUGOUDEBINGDUDEMUDE。ZHEIYIDIAN,MANBINGDUKENDINGMEIFAZUODAO,MUQIANMANBINGDUNENGDADAODEZUIGAOYOUXIAODIDUYIBANSHI108PFU/ml,DANSHIZHUSHEYIZHIXIAOSHUSUOXUDEBINGDULIANGZHISHAO1x108PFU/ml,ERYIZHIDASHUZHISHAOXUYAO109PFU/ml,ZHEIYANGBAOZHUANGYICIDEMANBINGDUZHINENGYONGYUYIZHIDONGWU,TONGSHI,YONGMANBINGDUDEHUA,YAODAYUE1mlDEZHUSHETIJICAINENGDADAODONGWUSUOXUDEBINGDULIANG,ZHEIGETIJIDEYETILIANGDUIYUDONGWUTISHIYOUHAIDE,YOUQISHIZHUSHENAOBU,DONGWUSHICHENGSHOUBUZHUDE,DANSHIYONGXIANBINGDUJIUBUHUICHUXIANZHEIYANGDEQINGKUANG,XIANBINGDUDEDIDUKEDADAO1011PFU/ml,YONGYUDONGWUSHIYANSHI,MEIZHIXIAOSHUZHIXUYAO1ul,ERMEIZHIDASHU10ulZHEIYANGJIKE,YICIBAOZHUANGDEXIANBINGDUZUYIWANCHENGYICIDONGWUSHIYANYANJIU,TONGSHIXIANBINGDUKEYIKUOZENGDE,YICIBAOZHUANGDEXIANBINGDUYONGWANHOUBUXUYAOZHONGTOUZAICIBAOZHUANG,ERZHIYAOZAIKUOZENGCHUNHUAJIKE,ZHEIYANGZAIJINGJICHENGBENHESHIJIANCHENGBENSHANGDOUHUIJIESHENGHENDUO。

WEILEGENGHAODETUCHUXIANBINGDUZAICHENGBENSHANGBIMANBINGDUDEYOUSHI,WOMENYONGLIUKONGBANHUANSUANFAKEYIGENGZHIGUANDEKANCHUXIANBINGDUYONGYUSHIYANYANJIUZAIJINGJICHENGBENSHANGDEYOUSHI。

YIXIAWEISHUOMING(GUOBIAODAQINGKUANG,YINWEIBINGDUKUDOUSHIGUOBIAODAKU)

HANHENGSHENGWUMANBINGDU/XIANBINGDUGANRANFUSHU(MOI)YUTIJIDUIYINGBIAO

GUIGE

DAYUESHUMU

MOIZHI

JIARUBINGDUSHULIANG

MANBINGDU

XIANBINGDU

96well

2-5×104

20-50

0.4-2.5×106

4-25ul

0.1-0.5ul

48well

1-2×105

20-50

0.2-1×107

20-100ul

0.5-2ul

24well

2-3×105

20-50

0.4-1.5×107

40-150ul

1-3ul

12well

5×105

20-50

1-2.5×107

100-250ul

2.5-5ul

6well

1-2×106

20-50

0.2-1×108

0.1-1ml

5-20ul

35mmdish

1-2×106

20-50

0.2-1×108

0.1-1ml

5-20ul

60mmdish

2-4×106

20-50

0.4-2×108

0.4-2ml

10-40ul

100mmdish

6-10×106

20-50

1.2-5×108

1.2-5ml

30-80ul

150mmdish

1-2×107

20-50

0.2-1×109

2-10ml

50-200ul

腺病毒滴度以1010/ml为准,其他滴度需相应换算;慢病毒滴度以108/ml为准,其他滴度需相应换算;
大约细胞数目是根据80%~100%细胞密度估算而出,具体细胞数请种细胞时进行细胞计数

 

 

FU:GEZHONGBINGDUZAITIGANRANMUDEXIBAOCAOZUOQUBIE

 

XIANBINGDU

MANBINGDU

NIZHUANLUBINGDU

GANRANFANGSHI

ZHIJIEJIARU

ZHIJIEJIARU

ZHIJIEJIARU

HUANYESHIJIAN

GANRANHOU6-8h

GANRANHOU24h

GANRANHOU24h

SHIFOUXUYAOSHAIXUAN

BUXUYAO,XIANBINGDUGANRANNENGLIHENQIANG

XUYAOSHAIXUANHUODEWENDINGGANRANZHU,MANBINGDUDEGANRANNENGLIBUQIANG

XUYAOSHAIXUANHUODEWENDINGGANRANZHU,NIZHUANLUBINGDUDEGANRANNENGLIBUQIANG

GUANCHASHIJIAN

RUDAIYOUGFPDENGYINGGUANGBIAOQIAN,24hHOUKEGUANCHADAOYINGGUANG,36hKEDADAOBIAODAGAOFENG,RUOBUDAIYOUYINGGUANGBIAOQIAN,ZEXUYAOJIANDING

DAIYOUGFPDENGYINGGUANGBIAOQIAN,24hHOUKEYIGUANCHADAOBIAODA,36hDADAOGAOFENG,RUOBUDAIYOUYINGGUANGBIAOQIAN,ZEBUKEZHIJIEGUANCHADAO,XUYAOJIANDING

DAIYOUGFPDENGYINGGUANGBIAOQIAN,24hHOUKEYIGUANCHADAOBIAODA,36hDADAOGAOFENG,RUOBUDAIYOUYINGGUANGBIAOQIAN,ZEBUKEZHIJIEGUANCHADAO,XUYAOJIANDING

SHIFOUXUYAOZHUZHUANJI

BUXUYAO

YOUSHIXUYAO,DANSHIGENJUJUTIQINGKUANGCAOZUO,YINWEIZHUZHUANJI,RUpolybrene,DUIXIBAODESHANGHAIBIJIAODA,YOUDEXIBAOKENENGCHENGSHOUBULE

YOUSHIXUYAO,DANSHIGENJUJUTIQINGKUANGCAOZUO,YINWEIZHUZHUANJI,RUpolybrene,DUIXIBAODESHANGHAIBIJIAODA,YOUDEXIBAOKENENGCHENGSHOUBULE

CHULEGENJUSHIYANYAOQIUHEMUDEJINXINGBUTONGBINGDUDEXUANZEWAI,DANSHIDUIYUTONGYIZHONGBINGDUWANGWANGHUIYOUDUOZHONGBINGDUBIAODAZAITI,YOUDEDAIYINGGUANG,YOUDEBUDAIYINGGUANG,YOUDEDAIGFPHUOZHERFP,YOUDEDAIluciferase,HAIYOUDESHITIAOJIANKONGZHIXINGZAITI,JUTIXUANZESHENMEYANGDEBINGDUZAITIYESHIYAOGENJUTEDINGDESHIYANMUDEJINXINGXUANZE,KEYIJUYIXIAJITIAOJINXINGXUANZE:

① XIANGEGFPZHEIYANGDEYINGGUANGDANBAITEYIXINGBUHAO,BUSHITEYIDEZAIMOUYILEIXIBAOZHONGBIAODA,DUIYUXIANGHUODETEDINGDEXIBAO,RUZUODONGWUZAITICHENGXIANGSHIYANSHI,DEDAODEXINHAOKENENGHUIBIJIAOLUAN,JIANYIBUXUANYONGDAIYOUEGFPYINGGUANGBIAOJIDEZAITI,KEYONGYONGQITABAOGAOJIYINRUluciferaseHUOZHERFP。

② EGFPDEJIFAGUANGBOZHANGWEI480nmZUOYOU,RUGUOSHIXIBAOSHIYAN,HOUXUSHIXIANGYAOZUOXIBAOZENGZHIHUOZHEDIAOWANGXIBAO,HOUMIANHUIXUYAOTONGGUOFITC (JIFAGUANGBOZHANGWEI488nm)JINXINGGUANCHA,ZHEIYANGHUIZAOCHENGCHONGTU,BURONGYIGUANCHA。LINGWAI,DANGHOUXUXIBAOSHIYANYAOZUODAOXUEYE/ZUZHIYEZHONGLDH JIANCE、PIRANHEDELIUSHIJIANCEXIBAOZHOUQISHIYAN、Ki67MIANYIYINGGUANGSHIYAN、BrduSHENRUSHIYAN、MTT/CCK8DENGXUYAOYONGDAOJIFAGUANGDESHIYAN,QIZHONGYONGDEBINGDUYESHIBUNENGYONGGFPBIAOJI,YINWEITAMENYONGDAODEJIFAGUANGBOZHANGHEGFPXIANGSI,ZHEIYANGKENENGHUIGANRAOSHIYANJIEGUO。

③ MIANYIYINGGUANGDENGSHIYANSHI,RUGUOYAOYANJIULIANGGEDANBAIDEXIANGHUZUOYONG,XUYAOLIANGZHONGYINGGUANG,ERDAIYOUYINGGUANGBIAOJIDEKANGTIHUIJIANSHAOLEYIGEKEYONGDEJIGUANGTONGDAO,ZHEIZHONGQINGKUANGXIA,ZHIKEYIQIZHONGYIGEBINGDUDAIYOUYINGGUANGBIAOJIHUOZHELIANGGEBINGDUDOUBUYAODAIYINGGUANGBIAOJI,ZHEIYANGFANGBIANHOUMIANSHIYONGDAIYOUYINGGUANGBIAOJIDEKANGTIJINXINGJIANCE。

④ YANJIULIANGZHONGDANBAIXIANGHUZUOYONG,HAIKEYIXUANZEYINGGUANGSUMEIBIAOJIZUOWEIBAOGAOJIYIN,KEYIXUANZELIANGZHONGBUTONGDEYINGGUANGSUMEI,HOUXUYONGSHENGWUXUEFANGFAJIANCEJISUANJIKE。

 

Chinese, Simplified
 
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