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海南七星彩 > 企业动态 > 转染是个坎,让我教你怎么翻过去 古朵生物√

企业动态

转染是个坎,让我教你怎么翻过去 古朵生物√
发布时间:2019-10-08   点击次数:147次

CONGJIYINBIAODADAORNAGANRAO,ZAIDAORUJINDAREDEJIYINZUBIANJI,TAMENDOULIBUKAIYIGEQIANTI:DNAHUORNAFENZINENGGOUDIDAORUXIBAONEIBINGFAHUIGONGNENG。RANER,WENTISHI,XIBAOBINGBUXIANGJIESHOUTAMEN,ERXIBAOMOYESHIYIGEKEPADEPINGZHANG,TADAIDESHIFUDIANHE,HESUANTONGYANGDAIFUDIANHE。

海南七星彩BUGUO,RENLEIDEZHIHUISHIWUXIANDE。RENMENYIJINGSHEJICHUGEZHONGFANGFALAIRAOGUOZHEIYIPINGZHANG,BAOKUOHUAXUEFANGFA,LIYONGDAIZHENGDIANDEZHUANRANSHIJILAIZHONGHEDAIFUDIANDEHESUAN;SHENGWUXUEFANGFA,CAIYONGJINGGUOYICHUANXUEGAIZAODEBINGDUJIANGFEIBINGDUJIYINZHUANYIZHIXIBAONEI;YIJIWULIFANGFA,ZAIXIBAOMOSHANGKAIKONG,JIANGHESUANZHIJIESONGRUXIBAO。

DANGRAN,GEZHONGXIBAOSHIYONGDEZHUANRANTIAOJIANDOUKENENGBUTONG,ZHENGSUOWEIJIAZHIMITANG、YIZHIpiSHUANG,BINGMEIYOUYIZHONGFANGZHISIHAIERJIEZHUNDEFANGFA。YINCI,YANJIURENYUANXUYAOGENJUNIDEXIBAOLEIXINGHESHIYANYAOQIULAIXUANZELIXIANGDEFANGFA。HEWEILIXIANGDEFANGFA?NEIJIUSHIZHUANRANXIAOLVGAO、XIBAODUXINGDI、DUIZHENGCHANGSHENGLIDEYINGXIANGzuiXIAO,BINGQIEJIANDANKEZHONGFU。ZHEIKANSITINGNAN,HAOZAIThermo Fisher(YUANLife Technologies)DAILAILEYIGEZHUANRANDEZHENGTIJIEJUEFANGAN,KEBANGZHUNIKUAISUSHIXIANMUBIAO。

JIYOUWENHEDEDNAZHUANRAN——Lipofectamine? 3000

 

海南七星彩DUIYUDNADEZHUANRAN,DAJIAWANGWANGZAISHIJIANXIANGDAOLipofectamine? 2000。ZHEIZHONGJINZHAOPAIDEYANGLIZIZHIZHITISHIJIJIHUSHIRENMENDESHOUxuan,MUQIANYINYONGWENXIANYIJINGCHAOGUO50,000PIAN。TONGGUODAIFUDIANHEDEHESUANYUDAIZHENGDIANHEDEHECHENGZHIZHITISHIJIZHIJIANDEJINGDIANZUOYONG,HESUAN-YANGLIZIZHIZHITISHIJIFUHEWUZIFAXINGCHENG。XIBAOTONGGUONEITUNZUOYONGSHEQUFUHEWU,RANHOUSHIFANGZHIBAOJIANG。YIDANJINRUXIBAO,ZHUANRANDNAZHUANYIZHIXIBAOHENEI,TONGGUOWEIZHIDEJIZHIBIAODA。

ZHISUOYIRUCISHOUHUANYING,SHOUXIANSHIYINWEITANENGGOUZHUANRANGEZHONGXIBAOXI,BANGZHUZAIGEZHONGSHIYANDEBUHUODEPIAOLIANGDEJIEGUO,QICIYEYINWEITADUIDNAHERNAJIEYOUXIAO,SHIHEDNAHERNADEGONGZHUANRAN。BUGUO,ZHUANRANXIAOLVHEXIBAODUXING,ZHENGRUYINGBIDEZHENGFANLIANGMIAN。Lipofectamine 2000ZAIHUODEZANYANGSHENGDETONGSHI,YESHOUDAODUXINGGAODEGOUBING。DUIYUWANQIANGDEXIBAOXIERYAN,ZHEIKENENGBUSHIWENTI,DANSHIJIAORUODEYUANDAIXIBAOHEGANXIBAOQUEZHETENGBUQI。

WEICI,Thermo FisherDEKEXUEJIAYOUHUALEZHUANRANGUOCHENGDESIGEBUZHOU,BINGJIEHEzuiXIANJINDEZHIZHITINAMIWEILIJISHU,KAIFACHUzuiXINDELipofectamine 3000SHIJI。TATIGONGLECHUZHONGDEZHUANRANXINGNENGHEKEZHONGFUDEJIEGUO,SHIYONGYUGEZHONGNANZHUANRANDEXIBAOHECHANGJIANDEXIBAO。YOUYUZHUANRANXINGNENGCHUZHONG,NIZAIZHUANRANGUOCHENGZHONGKEJIANSHAOSHIJIDEYONGLIANG,CONGERJINYIBUJIANGDIQIANZAIDEDUXINGFENGXIAN。

海南七星彩3000YU2000XIANGBI,ZHUANRANXIAOLVRUHE?YANJIURENYUANDANGRANYEJINXINGLEBIJIAO。TAMENSHIYONGZHEILIANGZHONGSHIJIZHUANRANLAIZIGEZHONGZUZHIDEAIXIBAO。YANJIUZHENGSHI,Lipofectamine 3000SHIJIDEZHUANRANXINGNENGMINGXIANGAOYU2000。SHIYONGLipofectamine 2000SHI,ZHIYOU25%DEAIZHENGXIBAOXIBEIRENWEISHIRONGYIZHUANRANDE(ZHUANRANXIAOLVGAOYU50%),ERSHIYONGLipofectamine 3000,ZHEIGEBILIGAODA60%。LISIBENDAXUEDERui Eduardo CastroBOSHIZAISHIYONGGUOLipofectamine 3000HOUXINGFENDISHUODAO:“WOMENFEICHANGGAOXINGQIEJINGYADIKANDAO, ZAINANYIZHUANRANDEXIBAOXIZHONGSHIYONGLipofectamine 3000,KEJIANGZHUANRANXIAOLVTIGAO10BEI,ERQIEJIANSHAOLEXIBAOSIWANGLV。JIEGUOZHENSHITAIBANGLE!”

YINCI,JISHISHINANZHUANRANDEXIBAO,SHIYONGLipofectamine 3000YEHENRONGYIHUODEMANYIDEJIEGUO。HAISHIYOUDIANBUMANYI?MEIGUANXI,Thermo FisherJIANYINICHANGSHIYIXIABUTONGDEDNANONGDUHEXIBAOMIDU。CAIYONGYANGLIZIZHIZHITIJIEDAODEZHUANRAN,ZHUANRANSHITIEBIXIBAODADAO70-90%HUIHE,XUANFUXIBAODA5 x 105ZHI2 x 106GEXIBAO/mL,KEYIHUODEzuiJIADEJIEGUO。HAISHIBUXING?NEIJIUSHISHImRNAZHUANRANBA。

mRNAYENENGZHUANRAN?DANGRAN!——Lipofectamine? MessengerMAX?

GUANYUmRNAZHUANRAN,RENMENWANGWANGYOUDUOZHONGWUJIE。YOURENRENWEI,mRNANANYIZHIBEI。QISHI,ZHEIFANGMIANDEGONGJUYOUHENDUO,BIRUThermo FisherDEmMESSAGE mMACHINE T7 UltraZHUANLUSHIJIHE,JIUNENGZHIBEI20-40 ?μgJIYONGXINGmRNA。HAIYOURENRENWEI,mRNABUWENDING,RONGYIJIANGJIE。No,no!ZHIYAOCAIQUYIXIEJIANDANDEYUFANGCUOSHI,JIUNENGQUEBAOmRNAWENDING:SHIYONGBUHANRNAMEIDESHIJIHEXITOU,FENZHUANGmRNABINGZHIYU-80?CBAOCUN。

YUDNAZHUANRANXIANGBI,mRNAZHUANRANZHIXUJINRUXIBAOZHI,ERFEIXIBAOHE,CONGERDADATIGAOLEYOUSIFENLIEHOUXIBAOHUOHUANMANFENLIEXIBAODEZHUANRANXIAOLV。ZHEIYANG,LIYONGLipofectamine MessengerMAXZHUANRANmRNAKEYISHIXIANGENGKUAISUDEDANBAIZHIBIAODA,ZAIZHUANRANXIBAOZHONGDADAOGENGGAODEBIAODAYIZHIXING,TEBIESHIHEYUANDAISHENJINGYUAN、SHENJINGGANXIBAOHEYONGSHENGHUASHENJINGXIBAO。

ZAIJIYINZUBIANJIDEYINGYONGZHONG,Lipofectamine MessengerMAXSHIJITONGGUOZHUANRANTIGAOLEGeneArt? CRISPRHESUANMEImRNADEJIANQIEHEZHONGZUXIAOLV,KEYIDADATIGAOJIYINXIUSHIDEXIAOLVBINGJIANHUAXIAYOUGUOCHENG。CIWAI,mRNABUHUIRUHE,ZHEIXIALAOBANZAIYEBUYONGDANXINJIYINZUZHENGHEDEFENGXIANLE。

YEXUNIHUIDANXIN,CAOZUOQILAIYIDINGHENFUZABA。QISHI,ZHIYAOZHIBEIHAOLEmRNA,ZHUANRANDEJIBENBUZHOUYUDNAXIANGTONG,BUWAIHUSHIXISHI、FUYUZHEIXIE,NANBUDAOZHUWEIGAOSHOU。KUANGQIEHAIKEYIGOUMAIYANGXINGDUIZHAOGFP mRNATONGSHIZHUANYIXIA,XIAOGUOHAOBUHAO,YIKANJIUZHIDAO。

CRISPRHAOBANGSHOU——Lipofectamine? CRISPRMAX? Cas9ZHUANRANSHIJI

海南七星彩DUIYUJIYINZUBIANJI,YOUXIERENSHIYONGCRISPRZHILI,YOUXIERENSHIYONGCas9 mRNA。QISHI,SHIYONGCas9DANBAIKEZAIYUANDAIXIBAOHEGANXIBAOZHONGHUODECHUSEDEQIEGEXIAOLV。TABIMIANLEZHUANLUHUOFANYI,KELIJIFAHUIZUOYONG,KUAISUQUCHU,zuidaCHENGDUDIJIANGDILETUOBAJIANQIEDEJILV。GeneArt? Platinum? Cas9HESUANMEIJIUSHIZHEIYANGYIZHONGYESHENGXINGDECas9DANBAI。

YEXUNIYAOWEN,RUHEJIANGCas9DANBAIDAORUXIBAONI?NEIJIUYAOYIKAOLipofectamine? CRISPRMAX? Cas9ZHUANRANSHIJILE。ZHEISHIKUANWEICRISPR-Cas9DANBAIZHUANRANERYOUHUADEZHIZHITINAMIWEILIZHUANRANSHIJI。ZHEIKUANSHIJIJIANDANYIYONG,DIDU,RUJINYIZAIiPSC、mESC、CHODENG20DUOZHONGXIBAOSHANGJINXINGLEJIANYAN。YUDIANCHUANKONGXIANGBI,TAGENGWEIWENHE,YINCISUOXUDEXIBAOJIAOSHAO,MEIGEFANYINGDECHENGBENJIAODI。CIWAI,TAYENENGQINGSONGKUOZHANDAOGAOTONGLIANGSHIYAN,SHICRISPRSHIYANDEHAOBANGSHOU。

海南七星彩RNAiDEBUERZHIXUAN——Lipofectamine? RNAiMAX

海南七星彩RNAiZUOWEIJIYINGONGNENGYANJIUDEJIBENGONGJU,RUJINYIYINGYONGZAIDANBAIZHIYIZHIYANJIU、BIAOXINGFENXI、TONGLUFENXIHEYAOWUBADIANFAXIANSHANG。XIAOFENZIRNA(siRNA、miRNA)YOUQIZISHENDETEDIAN,YEJIUXUYAOBUYIBANDEZHUANRANSHIJI。

海南七星彩Lipofectamine? RNAiMAX ZHUANRANSHIJIJIUSHIZHUANMENWEIXIAOFENZIRNAERYOUHUADE,KEYIDIJIANGXIAOFENZIRNADAORUGUANGFANDEXIBAOXI,BAOKUOCHANGJIANDEXIBAOLEIXING、GANXIBAO、YUANDAIXIBAOYIJINANZHUANRANDEXIBAO。

ZHEIZHONGZHUANRANSHIJIKEYIZAI10BEISHIJINONGDUFANWEINEIHUODEzuidaDEGANRAOXIAOGUOYIJIDEXIBAOCUNHUOLV,ZHEIGETEDIANSHIDELipofectamine? RNAiMAXDEZHUANRANTIAOJIANJIYIYOUHUA,YIBIANZAISHIYANZHONGSHIYONGzuiDIDEsiRNANONGDU,CONGERJIANGDIXIBAODUXING。JINGUANLipofectamine? RNAiMAXDESHUOMINGSHUJIANYIYI10 nM siRNA ZUOWEIYOUHUAGANRAOXIAOGUODEQISHINONGDU,DANSHIJINYONG1 nM siRNA YEKEYIHUODEJIAOHAODEGANRAOXIAOGUO。

海南七星彩Lipofectamine? RNAiMAXDECAOZUOYEXIANGDANGJIANDAN,ZHUANRANHOUWUXUQUCHUZHUANRANFUHEWU,WUXUGENGGAIHUOTIANJIAPEIYANGJI。RUGUOZHUANRANDEXIAOGUOBULIXIANG,KEYISHISHIFANXIANGZHUANRAN,HUOGAIBIANsiRNAYONGLIANG,HUOZHUANRANSHIDEXIBAOMIDU。LINGWAI,ZHEIKUANSHIJIZHINENGYONGYUXIAOFENZIRNADEZHUANRAN,BUNENGYONGYUGONGZHUANRAN。RUGUOXUYAOGONGZHUANRAN,ZHINENGJIEZHULipofectamine 3000HUO2000。

海南七星彩TINEIZHUANRANBUZAISHINANTI——Invivofectamine 3.0

ZHONGSUOZHOUZHI,YUANDAIXIBAODEZHUANRANBICHUANDAIXIBAOYAONANDEDUO,ERTINEIZHUANRANYOUBITIWAIZHUANRANYAONAN。DUIYUZHEIZHONGGAONANDUDESHIYAN,MIANQIANGSHIYONGCHUANTONGDETIWAIZHUANRANSHIJI(RUZHIZHITI)SHIMEIYOUXINGFUDE,zuiHAOHAISHIYONGZHUANMENDETINEIZHUANRANSHIJI,BIRUThermo FisherDEInvivofectamineXILIE。YIZHAYAN,InvivofectamineSHIJIYIJINGSHENGJIDAODISANDAI。

海南七星彩Invivofectamine 3.0SHIJISHIYIZHONGWUDONGWULAIYUANCHENGFENDEZHIZHINAMIKELI,TONGGUOWEIBUJINGMAIZHUSHE,NENGGOUJIANGsiRNAHEmiRNADAORUXIAOSHUGANZANGXIBAO。ZHUANRANXIAOGUORUHE?ZHUANJIAMENZUOGUOSHIYAN,JIANGInvivofectamine 3.0SHIJIYUZHENDUIFactor VIIHUOPPIBDEsiRNAHUNHE,YIMEIQIANKEXIAOSHUTIZHONG1 mgDEZHUSHELIANGJINXINGJINGMAIZHUSHE,zuiZHONGSHIXIANLEGAODA85%DEknockdown。YUZHIQIANDECHANPINXIANGBI,XINSHIJIZHIXUGENGSHAODEsiRNA,JIKESHIXIANYOUXIAOknockdown。CIWAI,DANCIZHUSHEInvivofectamine 3.0SHIJI/siRNAFUHEWUHOU,knockdownXIAOGUOKEWEICHIZHANGDA3ZHOU。XIANGYAOBAXIANGQITAQIGUAN?GUDUODEKEXUEJIAHAIJINXINGLEYIXIANHEPIZANGDESHIYAN,QUDELEHENHAODEXIAOGUO。

海南七星彩ZHIYUTADETINEIDUXING,ZHUANJIAMENYECENGPINGGUGUO。TAMENZAISHIJIZHUSHEHOUDEDUOGESHIJIANDIANJINXINGXUEYEHUAXUEFENXIJIXIBAOYINZIJIANCE。JIEGUOBIAOMINGZHUANRANHOUMEIGEGETIDESHENGWUBIAOZHIWUDESHUIPINGYUWEIZHUSHEGAISHIJIDEGETIXIANGBIBINGMEIYOUXIANZHUCHAYI。ZHEIBIAOMINGFUHEWUBIAOXIANCHUJIDIDETINEIDUXING。ZHEIZHONGSHIJIYEXIANGDANGWENDING,JISHIFANFUDONGRONG4CI,YEBUHUIZAOCHENGXINGNENGSUNSHI。

海南七星彩ZHONGjiWUQI——NeonZHUANRANXITONG

海南七星彩RUGUOZHEIXIEREMENGONGJUDOUBUGUANYONG,NEIWOMENZHINENGJICHUZHONGjiWUQI——NeonZHUANRANXITONG。ZHEISHIThermo FisherDIERDAIDETAISHIDIANCHUANKONGSHEBEI,KEJIANGHESUAN(BAOKUOZHILIDNAHEsiRNA)DAORUJIHURENYIYIZHONGDONGWUXIBAONEI,BAOKUONANYIZHUANRANDEYUANDAIXIBAO、GANXIBAOHEZAOXUEXITONGLAIYUANDEXIBAO,TONGSHIBAOCHIGAODEXIBAOHUOXING。

海南七星彩YUCHUANTONGDEDIANCHUANKONGYIBUTONG,NeonXITONGBINGMEIYOUCHUANSHUOZHONGDEDIANZHUANBEI,ERSHIYONGYIGETESHUDEYIYEQIXITOUQUDAI。YIYEQIXITOUNEISHIYONG24KDUJINDIANJI,KEXINGCHENGGENGWEIJUNYUNDEDIANCHANG,TONGSHIzuidaXIANDUDIJIANSHAOWENDUSHENGGAOHEpHZHIBIANHUAFUDU,ZAIQUEBAOXIBAOZHUANRANDETONGSHI,YEGENGJIAWENHE,BAOZHENGXIBAOCUNHUOLVGENGGAO。

NeonXITONGDAIYOULIANGZHONGXITOU:10 μlHE100 μl,RANGNIKEGENJUSHIYANYAOQIULINGHUOXUANZE,ERBUXIANGCHUANTONGDIANCHUANKONGYINEIYANGZHINENGZHUANRANGUDINGLIANGDEXIBAO。SHIYONGNeonXITONG,MEICIzuiSHAOKEZHUANRAN2×104GEXIBAO,zuiDUOKEZHUANRAN6×106GEXIBAO。DANGRAN,ZHEIBINGFEI。RUONIDEXIBAOGENGSHAOHUOGENGDUO,YEKEYISHI,JUZHUANJIAJIESHAO,YOUYIWEIYONGHUCENGCHENGGONGZHUANRANLE5000GEYUANDAImaoNANGXIBAO。

海南七星彩NeonYESHIYIGEKAIFANGDEXITONG,RANGYONGHUNENGGOUDANDUKONGZHIMEIGECANSHU。CHANGJIAYETIGONGLEYIGESHUJUKU,SHOUJIGEZHONGXIBAOLEIXINGDEJIANYISHEZHI,NIKEZAIZHEIGEJICHUSHANGYOUHUAHEDIAOZHENG。NeonNENGGOUDUIDUOZHONGNANYIZHUANRANDEXIBAOJINXINGZHUANRAN,BAOKUOYUANDAIXINGXINGJIAOZHIXIBAO、YUANDAISHUTUZHUANGXIBAO、Jurkat、XIAOSHUPEITAIGANXIBAODENG。DUOZHONGXIBAOLEIXINGKEHUODECHAOGUO75%DEZHUANRANXIAOLV。

 

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