RNA Yooj Yim Yooj Yim Cog Cov Ntaub Ntawv Cog Ntoo
Nta
■ Yooj yim thiab nrawm: Tag nrho RNA rho tawm los ntawm cov qauv cog tuaj yeem ua tiav hauv 30 feeb.
■ Muaj kev sib haum xeeb: Cov khoom siv no tsis yog tsuas yog haum rau cov qia thiab nplooj piv txwv, tab sis kuj rau polysaccharide/polyphenol-nplua nuj kuaj.
■ Muaj kev nyab xeeb thiab tsis muaj tshuaj lom: Cov tshuaj reagents lom xws li merc-mercaptoethanol, DTT, chloroform, phenol tsis xav tau.
Cov ntawv thov
■ Tsim nyog rau kev ua haujlwm nqes hav, suav nrog RT-PCR, RT-qPCR, kev sib xyaw ua ke, Sab Qaum Teb Blot, Dot Blot, PolyA tshuaj ntsuam, hauv kev txhais lus hauv vitro, RNase tiv thaiv kev tshuaj xyuas thiab cloning molecular, thiab lwm yam.
Txhua yam khoom tuaj yeem tsim kho rau ODM/OEM. Yog xav paub ntxiv,thov nyem Customized Service (ODM/OEM)
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Tag nrho RNA ntawm 65 mg nplooj ntawv piv txwv (nplooj clove, nplooj hibiscus, nplooj nplej, nplooj nplooj, ugli txiv hmab txiv ntoo nplooj, Prunus cerasifera nplooj, nplooj zaub, nplooj hnub, nplooj Kerria japonica nplooj), 150 mg kuaj cov kab mob (Lentinus edodes) thiab 200 mg kuaj cov paj ntoo (hnub-lily petals) tau muab rho tawm siv RNA Yooj Yim Yooj Yim Cog Cov Ntaub Ntawv Cuam Tshuam.Agilent Bioanalyzer 2100 txheeb xyuas qhov tshwm sim ntawm tag nrho RNA los ntawm Hnub-lily petals. |
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Agilent Bioanalyzer 2100 qhov kev tshuaj ntsuam tshwm sim ntawm tag nrho RNA los ntawm Hnub-lily nplaim paj. |
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RNA muab rho tawm los ntawm ntau qhov piv txwv teev tseg hauv lub rooj siv RNA Cov Khoom Siv Yooj Yim Yooj Yim Yooj Yim Cog. Kev tshem tawm ntim: 50 μl; Chaw thau khoom ntim: 2 μl. Cov electrophoresis tau ua ntawm 6 V/cm rau 15 feeb ntawm 1% agarose. |
A-1 Cell lysis lossis homogenization tsis txaus
---- Txo cov qauv siv, nce tus nqi ntawm lysis tsis, nce homogenization thiab lysis lub sijhawm.
A-2 Cov qauv piv txwv loj heev
---- Txo tus nqi siv piv txwv lossis nce tus nqi ntawm lysis tsis.
A-1 Tsis txaus lysis ntawm tes lossis homogenization
---- Txo cov qauv siv, nce tus nqi ntawm lysis tsis, nce homogenization thiab lysis lub sijhawm.
A-2 Cov qauv piv txwv loj heev
---- Thov xa mus rau lub peev xwm ua tiav siab tshaj plaws.
A-3 RNA tsis raug tshem tawm ntawm kab ntawv
---- Tom qab ntxiv RNase-Dawb dej, tso nws li ob peb feeb ua ntej centrifuging.
A-4 Ethanol hauv qhov tsim nyog
---- Tom qab yaug, cov centrifuge dua thiab tshem cov tshuaj ntxuav kom ntau li ntau tau.
A-5 Cell kab lis kev cai nruab nrab tsis raug tshem tawm kiag li
---- Thaum khaws cov cell, thov nco ntsoov tshem cov kab lis kev cai nruab nrab kom ntau li ntau tau.
A-6 Cov cell khaws cia hauv RNAstore tsis ua haujlwm zoo
---- RNAstore ntom ntau dua li qhov nruab nrab ntawm kab lis kev cai ntawm tes; yog li lub zog centrifugal yuav tsum tau nce. Nws tau pom zoo kom centrifuge ntawm 3000x g.
A-7 Cov ntsiab lus RNA qis thiab muaj ntau hauv cov qauv
---- Siv qhov piv txwv zoo los txiav txim siab tias qhov txiaj ntsig qis yog tshwm sim los ntawm cov qauv.
A-1 Cov khoom siv tsis tshiab
---- Cov ntaub so ntswg tshiab yuav tsum khaws cia hauv cov kua nitrogen tam sim lossis tam sim muab tso rau hauv RNAstore reagent kom ntseeg tau tias cov txiaj ntsig rho tawm.
A-2 Cov qauv piv txwv loj heev
---- Txo tus nqi qis.
A-3 RNase kev sib kisn
---- Txawm hais tias qhov tsis nyob hauv cov khoom siv tsis muaj RNase, nws yooj yim rau kis RNase thaum lub sijhawm txheej txheem rho tawm thiab yuav tsum tau ua nrog kev saib xyuas.
A-4 Electrophoresis paug
---- Hloov cov electrophoresis tsis thiab ua kom ntseeg tau tias cov khoom siv thiab Loading Tsis pub dawb ntawm RNase paug.
A-5 Kev thauj khoom ntau dhau rau electrophoresis
---- Txo tus nqi ntawm cov qauv thauj khoom, kev thauj khoom ntawm txhua lub qhov dej yuav tsum tsis pub tshaj 2 μg.
A-1 Tus nqi piv txwv loj heev
---- Txo tus nqi qis.
A-2 Qee qhov piv txwv muaj cov ntsiab lus DNA siab thiab tuaj yeem kho nrog DNase.
---- Ua RNase-DNase Kho DNase kom tau txais RNA daws, thiab RNA tuaj yeem siv ncaj qha rau kev sim tom ntej tom qab kho, lossis tuaj yeem ua kom huv ntxiv los ntawm RNA cov khoom siv ua kom huv.
Rau cov iav, ci ntawm 150 ° C rau 4 h. Rau cov ntim yas, raus rau hauv 0.5 M NaOH rau 10 feeb, tom qab ntawd yaug kom huv nrog RNase tsis pub dej thiab tom qab ntawd ua kom tsis huv kom tshem tawm RNase. Cov tshuaj reagents lossis cov kev daws teeb meem siv hauv kev sim, tshwj xeeb tshaj yog dej, yuav tsum tsis pub dawb ntawm RNase. Siv RNase-dawb dej rau txhua qhov kev npaj ua tshuaj reagent (ntxiv dej rau hauv lub khob iav huv, ntxiv DEPC mus rau qhov kawg ntawm 0.1% (V/V), co thaum hmo ntuj thiab autoclave).
Pawg khoom
Vim Li Cas Xaiv Peb
Txij li nws tau tsim, peb lub Hoobkas tau tsim thawj cov khoom lag luam hauv ntiaj teb nrog ua raws txoj cai
zoo ua ntej. Peb cov khoom tau txais txiaj ntsig zoo hauv kev lag luam thiab muaj txiaj ntsig zoo ntawm cov neeg siv khoom qub thiab qub ..
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