Cov Ntawv Sib Nqus/Xovtooj/Ntshav Tag Nrho Cov Khoom RNA
Nta
■ Yooj yim thiab nrawm: Ultrapure tag nrho RNA tuaj yeem tau txais hauv 60 feeb.
■ Kev nkag siab siab: Nws tuaj yeem ua tau raws li qhov xav tau ntawm phau ntawv rho tawm nrog rau kev rho tawm ntawm ntau yam kev sib tw siab.
■ Muaj kev nyab xeeb thiab tsis muaj tshuaj lom: Tsis xav tau tshuaj reagent xws li phenol/chloroform.
Kev qhia tshwj xeeb
Hom: Sib nqus hlaws hom rho tawm.
Qauv: Tissues, cells and blood.
Lub Hom Phiaj: Tag Nrho RNA
Pib ntim: 5-20 mg, tsis pub tshaj 1 × 107, 0.1-1.5 ml
Lub sijhawm ua haujlwm: 60 min
Cov ntawv thov nqes hav: RT-PCR/RT-qPCR, kev tsim tsev qiv ntawv NGS, 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 tau muab rho tawm los ntawm 20 mg nas lub siab nrog TIANGEN Sib nqus Tissue/Cell/Ntshav Tag nrho RNA Cov Khoom thiab cov khoom cuam tshuam los ntawm lwm tus neeg muag khoom. 5 μl ntawm 200 μl eluate tau thauj mus rau 1% agarose gel electrophoresis, 6 V/cm. Xaus: Kev rho tawm cov txiaj ntsig, kev coj dawb huv thiab ruaj khov ntawm TIANGEN Sib Nqus Tissue/Cell/Ntshav Tag Nrho RNA Cov Khoom tau zoo dua li ntawm lwm tus neeg muag khoom. |
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Tag nrho RNA tau muab rho tawm los ntawm 20 mg ntawm lub raum nrog TIANGEN Sib Nqus Tissue/Cell/Ntshav Tag nrho RNA Cov Khoom hauv TIANGEN TGuide S32 lossis Thermo Kingfisher Flex96 feem. 5 μl ntawm 200 μl eluate tau thauj mus rau 1% agarose gel electrophoresis, 6 V/cm. Cim: TIANGEN D15000 DNA Marker. Xaus: Daim Ntawv Sib Nqus Sib Nqus/Cell/Ntshav Tag Nrho RNA Cov Khoom muaj cov txiaj ntsig zoo rho tawm thiab kev coj dawb huv hauv cov tshuab rho tawm sib txawv. |
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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