qPCR背景介绍
ZAISHISHIYINGGUANGDINGLIANGPCRZHONG,DUIMEIYILUNXUNHUANDEYINGGUANGXINHAOQIANGDUJINXINGJIANCEBINGJILU,SUIZHEFANYINGDEJINXING,JIANCEDAODEYINGGUANGXINHAOKEYIHUIZHICHENGYITIAOQUXIAN,JIWEIKUOZENGQUXIAN。YIBANERYAN,KUOZENGQUXIANKEYIFENCHENGSANGEJIEDUAN:JIXIANQI(Baseline phase),ZHISHUZENGZHANGQI(Log-linear phase)HEPINGTAIQI(Plateau phase)(TUYI)。ZAIKUOZENGZAOQIJIJIXIANQI,KUOZENGDEYINGGUANGXINHAOBEIYINGGUANGBEIJINGXINHAOSUOYANGAI,WUFAPANDUANCHANWULIANGDEBIANHUA。ZAIPINGTAIQI,KUOZENGCHANWUYIBUZAICHENGZHISHUJIDEZENGJIA,PCRDEZHONGCHANWULIANGYUQISHIMOBANLIANGZHIJIANWUXIANXINGGUANXI,SUOYIGENJUZUIZHONGDEPCRCHANWULIANGBUNENGJISUANCHUCHUSHIMOBANLIANG。ZHIYOUZAIYINGGUANGXINHAOZHISHUKUOZENGJIEDUAN,PCRCHANWULIANGDEDUISHUZHIYUQISHIMOBANLIANGZHIJIANCUNZAIXIANXINGGUANXI,KEZAICIJIEDUANJINXINGDINGLIANGFENXI。
流程图
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HANHENGSHENGWUTIANBODIANJING(SHANGHAI)YOUXIANTIANBODIANJINGTIGONGCONGYINWUSHEJI,XIBAOHUOZHEZUZHIYANGPINCHULI,qPCRJIANCEYIJIWANZHENGDESHUJUBAOGAODENGFUWU。WEININTIGONGZUIYOUZHIDEFUWU,WEININDEKEYANBAOJIAHUHANG。
KEHUTIGONG:
1.细胞:细胞数106-107;组织:总量>100mg;DNA或RNA样品:总量>2 μg
2.BAJIYINXINXI:BAOKUOJIYINXULIEHUOGenBank DENGLUHAODENG
3.BEIJINGZILIAO:BAOKUOSHENGWUWUZHONGXINXI、JIYINGONGNENG、YONGTU、TEXING、DNA/RNA LAIYUANDENG
(BEIZHU:QINGZAISONGYANGQIAN,YUJISHUQUERENJUTISONGYANGQINGKUANG。)
实例展示:
(Hou Z ,Qin X ,Hu Y ,et al. 2019)
ZAIYANJIUZHANGQIDUANLIANBAOHUXINZANGDEFENZIJIZHIZHONG,YANJIURENYUANDUIYUNDONGDASHU(Exe-exo)HEJIUZUODASHU(Sed-exo)DEWAIMITIJINXINGmiRNA-seqHEqPCR,LAIQUEDINGYANJIUMUBIAOYUXINZANGBAOHUXIANGGUANDEmiR-342 5p。
CANKAOWENXIAN:
Hawkins, S. F. C, Guest, P. C. Multiplex Analyses Using Real-Time Quantitative PCR[J]. Methods in Molecular Biology,2016,1546:125-133.
Wong M L , Medrano J F . Real-time PCR for mRNA quantitation[J]. BioTechniques, 2005, 39(1):75-85
Hou Z , Qin X , Hu Y , et al. Longterm Exercise-Derived Exosomal miR-342-5p: A Novel Exerkine for Cardioprotection[J]. Circulation Research, 2019,124(9):1386-1400.
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