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技术文章
非接触式超声波破碎仪在染色质免疫沉淀技术中的应用

宁夏11选5走势图    FEIJIECHUSHICHAOSHENGBOPOSUIYIZHUYAOYONGYUWUJUNPOSUIXIBAO,GEZHELIXINGUANNENGDADUANRANSETI,KEZAIMIFENGRONGQIXIAJINXINGPOSUI,CHAOSHENGBOTANTOUYUYANGPINBUJIECHU,BIMIANJIAOCHAWURAN。KEXUANPEILENGQUESHUIXUNHUANXITONG,BIANYUYANGPINZAI4℃SHUIYUCHAOSHENG,NENGLIANGFENBUJUNYUN,CHAOSHENGZUOYONGWANQUAN,SHIYANZHONGFUXINGHAO,JIEGUOKEKAOXINGGAO。

    ZHENHESHENGWUDEJIYINZUDNAYIRANSEZHIDEXINGSHICUNZAI。YINCI,YANJIUDANBAIZHIYUDNAZAIRANSEZHIHUANJINGXIADEXIANGHUZUOYONGSHICHANMINGZHENHESHENGWUJIYINBIAODAJIZHIDEJIBENTUJING。RANSEZHIMIANYICHENDIANJISHU(chromatin immunoprecipitation assay, CHIP )SHIMUQIANYANJIUTINEIDNAYUDANBAIZHIXIANGHUZUOYONGDEFANGFA。

宁夏11选5走势图    RANSEZHIMIANYIGONGCHENDIAN(CHIP)DEYUANLI:

宁夏11选5走势图    SHIZAIHUOXIBAOZHUANGTAIXIAGUDINGDANBAIZHI-DNAFUHEWU,BINGJIANGQISUIJIQIEDUANWEIYIDINGZHANGDUFANWEINEIDERANSEZHIXIAOPIANDUAN,RANHOUTONGGUOMIANYIXUEFANGFACHENDIANCIFUHETI,TEYIXINGDIFUJIMUDEDANBAIJIEHEDEDNAPIANDUAN,TONGGUODUIMUDEPIANDUANDECHUNHUAYUJIANCE,CONGERHUODEDANBAIZHIYUDNAXIANGHUZUOYONGDEXINXI。

   CHIPSHIYANBUZHOU:

1、XIBAOGUDING

   JIAQUANNENGYOUXIAODESHIDANBAIZHI-DANBAIZHI,DANBAIZHI-DNA,DANBAIZHI-RNAJIAOLIAN,XINGCHENGSHENGWUFUHETI,FANGZHIXIBAONEIZUFENDEZHONGXINFENBU。JIAOLIANSUOYONGDEJIAQUANZHONGNONGDUWEI1%,JIAOLIANSHIJIANTONGCHANGWEI5FENZHONGDAO1GEXIAOSHI。XUZHUYIDESHI,JIAOLIANSHIJIANRUGUOGUOZHANG,XIBAORANSEZHINANYIYONGCHAOSHENGBOPOSUI,YINGXIANGChIPJIEGUO,ERQIESHIYANCAILIAOYERONGYIZAILIXINGUOCHENGZHONGDIUSHI。JIAOLIANSHIJIANRUGUOGUODUAN,ZEJIAOLIANBUWANQUAN,CHANSHENGJIAYINXING。JIAQUANDEJIAOLIANFANYINGKEBEIJIARUDEGANANSUANZHONGZHI。

2、RANSEZHIPIANDUANHUA

   交联后的染色质需被超声波切成400~600bp的片段,以便目的蛋白的暴露,利于抗体识别,其片段破碎的均匀一致性对结果影响至关重要。传统的超声波破碎是利用机械力断裂染色质,容易引起升温或产生泡沫,这都会引起蛋白质变性,进而影响ChIP的效率。而非接触式超声波破碎仪采用温和的破碎方式,保证了蛋白的活性,DNA破碎效果均一,同时外接水循环仪保证了实验的温度稳定性,成为目前ChIP实验的优先考虑设备。

3、RANSEZHIMIANYICHENDIANFANYING

InputDUIZHAO:

ZAIJINXINGMIANYICHENDIANQIAN,XUYAOQUYIBUFENDUANLIEHOUDERANSEZHIZUOInputDUIZHAO。InputXUYAOYUCHENDIANHOUDEYANGPINDNAYIQIJINGGUONIZHUANJIAOLIAN,DNACHUNHUA,YIJIzuiHOUDEJIANCE。InputDUIZHAOBUJINKEYIYANZHENGRANSEZHIDUANLIEDEXIAOGUO,HAIKEYIGENJUInputZHONGDEBAXULIEDEHANLIANGYIJIRANSEZHICHENDIANZHONGDEJIANYIBAXULIEDEHANLIANG,ANZHAOQUYANGBILIHUANSUANCHUChIPDEXIAOLV,SUOYIInputDUIZHAOSHIChIPSHIYANBIBUKESHAODEBUZHOU。

BeadsXUANZE:

宁夏11选5走势图JIEXIALAI,LIYONGMUDEDANBAIZHIDETEYIKANGTITONGGUOKANGYUAN-KANGTITEYIFANYINGXINGCHENGDNA-DANBAIZHI-KANGTIFUHEWU,RANHOUSHIYONGAgarose beadsHUOMagnabeadsCHENDIANCIFUHEWU,TEYIXINGDIFUJIYUMUDEDANBAIJIEHEDEDNAPIANDUAN。ZAIJINGGUODUOCIXIDI,CHUQUFEITEYIJIEHEDERANSEZHIHOU,YONGSDS+NaHCO3XITUOMIANYICHENDIANFUHEWU。

KANGTIXUANZE:

RANSEZHIMIANYICHENDIANSUOXUANZEDEMUDEDANBAIDEKANGTISHIChIPSHIYANCHENGGONGDEGUANJIAN。YINWEIZAIDANBAIZHIYURANSEZHIJIAOLIANJIEHESHI,KANGTIDEKANGYUANBIAOWEIKENENGYINWEIYUJIEHEWEIDIANDEJULITAIJIN,BUNENGBEIKANGTISHIBIE,SUOYIBUNENGYOUXIAODIZAITINEIXINGCHENGMIANYICHENDIANFUHEWU,ZHIJIEYINGXIANGChIPDEJIEGUO。

YINXINGDUIZHAOSHEZHI:

宁夏11选5走势图ZAIZUOChIPSHIYANSHI,XUSHEZHIDUIZHAO,YIBIANYUDUISHIYANJIEGUODEKEKAOXINGJINXINGPINGGU。YANGXINGKANGTIHEYINXINGKANGTIDUIZHAOSHIJIBENDESHIYANDUIZHAO。YANGXINGKANGTITONGCHANGXUANZEYUYIZHIXULIEXIANGJIEHEDEBIJIAOBAOSHOUDEDANBAIDEKANGTI,CHANGYONGDEBAOKUOZUDANBAIKANGTIHUORNAPolymerase IIKANGTIDENG。YINXINGKANGTITONGCHANGXUANZEMUDEDANBAIKANGTISUZHUDEIgGHUOXUEQING。MUDEDANBAIKANGTIDEJIEGUOYUYANGXINGKANGTIHEYINXINGKANGTIDEJIEGUOXIANGBIJIAO,CAINENGDECHUZHENGQUEJIELUN。LINGWAI,HAIYINGKAOLVMUDEDANBAIKANGTIYUDNADEFEITEYIXINGJIEHEDEKENENG,SUOYITONGCHANGHAIHUIXUANZEYIDUIYINXINGYINWU,JIMUDEDANBAIKENDINGBUHUIJIEHEDEDNAXULIE,ZUOWEIGAIKANGTIDEYINXINGDUIZHAO。JIAOYOUDEYINXINGDUIZHAOYINWUSHIZAIBAXULIESHANGYOUDEYIDUANYUMUDEDANBAIKENDINGBUNENGJIEHEDEXULIE。RUGUOMUDEDANBAIMEIYOUSHANGPINHUADESHIYONGYURANSEZHIMIANYICHENDIANSHIYANDEKANGTI,ZHIYOUQITAYONGTUDEKANGTISHI,KEYIXIANZUODANBAIZHIMIANYICHENDIANJIANCE。RUGUOKANGTIKEYICHENGGONGDECHENDIANDANBAI,ZAIJINXINGRANSEZHIMIANYICHENDIANSHIYANDEJIANCE。

4、JIAOLIANFANYINGDENIZHUANHEDNADECHUNHUA

   YONGBUHANDNaseDERNaseHEProteinase K,65℃BAOWEN6XIAOSHINIZHUANJIAOLIAN,JINGDNACHUNHUAZHUHUISHOUDNACHOUTI、YICHUNCHENDIANCHUNHUADNA。ZAINIZHUANJIAOLIANSHIBUSHIYONGProteinase K,RANHOUYONGBINGTONGHUISHOUYOUJIXIANGZHONGDEDANBAIZHI,JINXINGFENXI。

5、DNADEJIANDING

   CHANGYONGDEDNADEJIANDINGFANGFASHIBANDINGLIANGPCRHEReal-time PCR。YOUYUQIDONGZIQUYUXULIEDUOYANGXINGDETEDIAN,SUOYIBUTONGDEXIBAOXIHUOBUTONGDEDONGWUPINXIDETONGYIJIYINDEQIDONGZIXULIEYOUKENENGBUTONG,YINCIKESHEJIDUODUIYINWULAIFANFUYANZHENGChIPSHIYANDEJIEGUO。

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