Small RNA测序

产品介绍

皇朝国际small RNACEXUSHISHENGMINGHUODONGZHONGYAODEDIAOKONGYINZI,ZAIJIYINBIAODADIAOKONG、SHENGWUGETIFAYU、DAIXIEJIJIBINGDEFASHENGDENGSHENGLIGUOCHENGZHONGQIZHEZHONGYAODEZUOYONG。IlluminaNENGGOUDUIXIBAOHUOZHEZUZHIZHONGDEQUANBUSmall RNAJINXINGSHENDUCEXUJIDINGLIANGFENXIDENGYANJIU。

miRNA鉴定

miRNAZHUANLUQISHIWEIDIANDUOWEIYUJIYINJIANGEQU、NEIHANZIYIJIBIANMAXULIEDEFANXIANGHUBUXULIESHANG,QIQIANTIJUYOUBIAOZHIXINGDEFAJIAJIEGOU,CHENGSHUTIDEXINGCHENGSHIYOUDicer/DCLMEIDEJIANQIESHIXIANDE。DUIYUYIZHImiRNADEJIANDING,WOMENJIANGBIDUISHANGCANKAOJIYINZUDEreadsXULIEYUYIZHImiRNASHUJUKUmiRBase(v21)ZHONGDECHENGSHUmiRNAXULIEJINXINGBIDUI。ZHENDUImiRNADESHENGWUTEZHENG,DUIYUWEIJIANDINGDAOYIZHImiRNADEXULIE,BAIMAIKELIYONGmiRDeep2RUANJIANJINXINGXINmiRNADEYUCE。

mirna-2
mirna-3

基因表达水平分析

LIYONGZHUANLUZUSHUJUJIANCEJIYINBIAODAJUYOUJIAOGAODELINGMINDU。TONGGUOFPKMMIDUTUHEXIANGXIANTUBUJINKEYIFANYINGDANGEYANGPINJIYINBIAODASHUIPINGFENBUHELISANCHENGDU,HAIKEYIZHIGUANDEBIJIAOBUTONGYANGPINDEZHENGTIJIYINBIAODASHUIPINGCHAYI。

差异表达miRNA聚类分析

JULEIFENXIYONGYUPANDUANCHAYIJIYINZAIBUTONGSHIYANTIAOJIANXIADEBIAODAMOSHI,KETONGGUOJIANGBIAODAMOSHIXIANGTONGHUOXIANGJINDEJIYINJUJICHENGLEI,CONGERSHIBIEWEIZHIJIYINDEGONGNENGHUOYIZHIJIYINDEWEIZHIGONGNENG,TONGLEIJIYINKENENGJUYOUXIANGSIDEGONGNENGHUOGONGTONGCANYUTONGYIDAIXIEGUOCHENG。DUISHAIXUANCHUDECHAYIBIAODAmiRNAZUOCENGCIJULEIFENXI,JIANGJUYOUXIANGTONGHUOXIANGSIBIAODAXINGWEIDEmiRNAJINXINGJULEI。

mirna-4
mirna-5

差异miRNA靶基因注释

SHENGWUTINEI,ZAITEDINGTIAOJIANXIAMOUXIEJIYINDUIYINGDEZHUANLUBENHUIXINGCHENGmiRNAQIANTI,SHIQUBIANMAGONGNENG,CONGERYINGXIANGSHENGWUXUEBIAOXING。YINCI,ZHENDUImiRNADEBAJIYINFENBIEJINXINGGOHEKEGGZHUSHIJIFUJIFENXI,YOUZHUYUSHENRUWAJUEXIAORNAGONGNENG。

Q1. 动植物在miRNA预测中有什么差别:

答:植物miRNA可与mRNA的编码区完全互补配对,并通过诱导mRNA降解而发挥抑制表达的作用,我们可以直接通过比对来筛选靶基因
动物miRNA则可与mRNA的3’UTR区部分互补配对结合,进而抑制翻译的进行,一般的,miRNA与mRNA的配对区域位于miRNA的5’端的 2-8个碱基,称为种子区,只要种子区能与mRNA互补配对即可发挥作用,这也是一个miNRA能够调控数百条mRNA的原因。
①对于动物预测靶基因我们不是直接通过比对进行筛选的。
皇朝国际 ②在动物预测时,由于我们没法给出3‘utr区,因此我们截取mRNA5’端前2000bp使用动物预测靶基因方法进行预测,target_length指的就是我们截取该基因的长度。

Q2. 分析结果中如何确定miRNA对应的pre-miRNA的数量(别人论文中有这样的结论:We identified 83 potential novel miRNAs, which corresponded to 95 genomic loci.)?

DA:WENXIANZHONGDEMIAOSHUSHIZHIMOUXIECHENGSHUDEmiRNAXULIEKENENGLAIZIYUBUTONGDEmiRNAQIANTIXULIE,WUCANXIANGMUZHONGFENXIDEnovel miRNASHIZHENDUIMEIGEUnigeneFENXIDE,MINGMINGYESHIYUUnigeneDUIYING,MEIYOUDUICHENGSHUmiRNADEXULIEJINXINGZHONGFUDEFENXI。DUICHENGSHUDEmiRNADEXULIEJINXINGQUZHONGFUTONGJI,LIRUMOUXIANGMUYUCECHULE229GEmiRNA,QIZHONGYOU6GEmiRNAFENBIELAIZIYULIANGGEBUTONGDEQIANTIXULIE,GUKEBIAOSHUWEI:YUCECHULE223GEmiRNA,DUIYINGLE229GEQIANTIXULIE。

Q3. 目前对于小RNA靶基因的结果验证方法有哪些?

DA:

  1. RT-PCR对miRNA进行定量验证;
  2. miRNA过表达;
  3. 荧光素酶法,过表达miRNA测定靶基因表达量;把靶基因连上一个荧光素酶,如果靶基因表达量下降,可检测到的荧光信号则减弱
  4. 降解组测序
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