Tsiaj Tag Nrho RNA Isolation Cov Khoom Siv Tag Nrho RNA Extraction thiab Purification Cov Khoom Siv rau Tsiaj Tsiaj & Cell
Kit Description:
Ceev ceev thiab ua tau zoo extract high-purity thiab high-zoo tag nrho RNA los ntawm ntau yam tsiaj cov ntaub so ntswg.
Tsis tas yuav txhawj txog RNA degradation.Tag nrho cov kab ke yog RNase-Free
Kev tshem tawm DNA zoo siv DNA-Tshuaj Kem
Tshem tawm DNA yam tsis tau ntxiv DNase
Yooj yim-tag nrho cov haujlwm ua tiav ntawm chav tsev kub
Fast-kev ua haujlwm tuaj yeem ua tiav hauv 30 feeb
Kev nyab xeeb - tsis muaj cov organic reagent siv
Siab purity-OD260/280≈1.8-2.1
Cov khoom piav qhia
50 Preps, 200 Preps
Cov khoom siv no siv lubspin kem thiab formulatsim los ntawm peb lub tuam txhab, uas muaj peev xwm extract high-purity thiab high-zoo tag nrho RNA los ntawm ntau yam tsiaj cov ntaub so ntswg nrog high efficiency.It muab ib tug npaum DNA-Tshuaj Kem, uas tau yooj yim cais thiab adsorb genomic DNA los ntawm supernatant thiab cov ntaub so ntswg lysate, yooj yim thiab txuag sij hawm;RNA nkaus xwb Kem tuaj yeem khi RNA zoo thiab tuaj yeem ua tiav ib txhij nrog cov qauv tshwj xeeb Ntau cov qauv.
Tag nrho cov kab ke yog RNase-Free, kom cov RNA rho tawm tsis degraded;Buffer RW1, Buffer RW2 tsis siv lub tshuab ntxhua khaub ncaws, kom cov RNA tau txais tsis muaj protein ntau, DNA, ion, thiab organic compound pollution.
Cov khoom siv
| Tsiaj Tag Nrho RNA Isolation Kit | ||
| Cov khoom siv | TSI-03011 | TSI-03014 |
| 50 T | 200 T | |
| Tsis muaj RL1 * | 25ml ib | 100 ml |
| Tsis muaj RL2 | 15ml ib | 60ml ib |
| Buffer RW1* | 25ml ib | 100 ml |
| Buffer RW2 | 24ml ib | ib 96ml |
| RNase-Dawb ddH2O | 10 ml | 40 ml |
| RNA nkaus xwb | 50 | 200 |
| DNA-Tswj Kem | 50 | 200 |
| Phau ntawv qhia | 1 qob | 1 qob |
Cov ntaub ntawv khoom
| Hom ntawv | Khiav kab | Purification tivthaiv | Foregene kem, reagent |
| Flux | 1-24 qauv | Sijhawm rau kev npaj | ~ 30 feeb (24 kuaj) |
| Centrifuge | Lub rooj centrifuge | Pyrolysis sib cais | Kev sib cais centrifugal |
| Qauv | Tsiaj cov ntaub so ntswg;cell | Cov qauv nqi | Cov ntaub so ntswg: 10-20 mg;Cell: (1-5) × 106 |
| Elution ntim | 50-200 hli | Max loading ntim | 850l ua |
Nta & qhov zoo
■ Tsis tas yuav txhawj txog RNA degradation;tag nrho cov kab ke yog RNase-Free
■ Tshem tawm DNA-siv DNA-Tshuaj Kem
■ Tshem tawm DNA yam tsis tau ntxiv DNase
■ Kev ua haujlwm yooj yim-txhua yam ua tiav ntawm chav tsev kub
■ Ceev -kev ua haujlwm tuaj yeem ua tiav hauv 30 feeb
■ Kev nyab xeeb-tsis muaj cov organic reagent yuav tsum tau
■ High purity -OD260/280≈1.8-2.1
Daim ntawv thov Kit
Nws yog qhov tsim nyog rau kev rho tawm thiab ua kom huv ntawm tag nrho RNA los ntawm ntau yam tshiab lossis khov tsiaj cov ntaub so ntswg lossis kab lis kev cai.
Khoom parameters
■ Cov ntawv thov hauv qab: thawj-strand cDNA synthesis, RT-PCR, molecular cloning, Northern Blot, thiab lwm yam.
■ Cov qauv: cov ntaub so ntswg tsiaj, cov kab lis kev cai
■ Siv: Cov ntaub so ntswg 10-20mg, Cells (2-5) × 106
■ Qhov siab tshaj plaws DNA khi muaj peev xwm ntawm purification kem: 80 μg
■ Elution ntim: 50-200 μl
Daim duab
Animal Total RNA Isolation Kit kho 20mg
Cov qauv nas tshiab, noj 5% purified tag nrho RNA 1% agar
Glycogel electrophoresis
1: Spleen 2: Lub raum
3: Lub siab 4: Lub siab
Cia thiab txee lub neej
Cov khoom siv tuaj yeem khaws cia rau 24 lub hlis ntawm chav tsev kub (15-25 ℃) lossis 2-8 ℃ rau lub sijhawm ntev.Buffer RL1 tuaj yeem khaws cia ntawm 4 ℃ rau 1 lub hlis tom qab ntxiv β-mercaptoethanol (optional).
Cov lus hais
1.PIB: 18.808.Zheng, Q., Qin, F., Luo, R., et al.mRNA-Loaded Lipid-zoo li nanoparticles rau lub siab Base Editing ntawm Optimization ntawm Central Composite Design.Adv.Ua haujlwm.Mater.2021, 31, 2011068.doi:10.1002/adfm.202011068.
2.PIB: 18.187.He X, Hong W, Yang J, et al.Spontaneous apoptosis ntawm cov hlwb nyob rau hauv kev kho mob qia cell npaj exert immunomodulatory teebmeem los ntawm kev tso tawm ntawm phosphatidylserine.Teeb liab Transduct Target Ther.2021 Lub Xya Hli 14; 6(1): 270.doi: 10.1038/s41392-021-00688-z.
3.Ib: 17.97: Dai Z, Liu H, Liao J, et al.N7-Methylguanosine tRNA kev hloov kho txhim kho oncogenic mRNA kev txhais lus thiab txhawb kev loj hlob ntawm intrahepatic cholangiocarcinoma.Mol Cell.2021 Jul 29:S1097-2765(21)00555-4.doi: 10.1016/j.molcel.2021.07.003.
4.Ib: 9.225: Cao X, Shu Y, Chen Y, et al.Mettl14-Mediated m6A Kev Hloov Kho Ua kom lub siab rov tsim dua tshiab los ntawm kev tuav cov Endoplasmic Reticulum Homeostasis.Cell Mol Gastroenterol Hepatol.2021; 12(2): 633-651.doi: 10.1016/j.jcmgh.2021.04.001.
RNA isoaltion cov khoom siv rau lwm yam qauvmuaj:
Cell, cog, kab mob, ntshav, thiab lwm yam.
RNA tsis raug rho tawm lossis RNA tawm los tsawg
Feem ntau muaj ntau yam uas cuam tshuam rau kev rov qab ua haujlwm zoo, xws li: cov ntaub so ntswg qauv RNA cov ntsiab lus, kev ua haujlwm, elution ntim, thiab lwm yam.
1. Cov dej khov da dej lossis cryogenic (4 ° C) centrifugation tau ua thaum lub sijhawm ua haujlwm.
Kev pom zoo: Ua haujlwm ntawm chav tsev kub (15-25 ° C) thoob plaws hauv tag nrho cov txheej txheem, tsis txhob dej khov thiab centrifuge ntawm qhov kub qis.
2. Cov qauv khaws cia tsis raug lossis ntau lub sijhawm khaws cia.
Kev pom zoo: Khaws cov qauv ntawm -80 ° C lossis khov hauv cov kua nitrogen thiab tsis txhob rov siv dua khov-thaw;sim siv cov ntaub so ntswg tshiab los yog kab lis kev cai rau RNA rho tawm.
3. Cov qauv lysis tsis txaus.
Daim ntawv pom zoo: Thaum homogenizing cov ntaub so ntswg, xyuas kom meej tias cov ntaub so ntswg txaus homogenized thiab cov ntaub so ntswg hlwb txaus txaus los piav qhia txog kev tso tawm RNA.
4. Lub eluent tsis ntxiv kom raug.
Kev Pom Zoo: Paub meej tias RNase-Free ddH2O yog ntxiv dropwise mus rau nruab nrab ntawm lub purification kem membrane.
5. Qhov tseeb ntim ntawm ethanol tsis tau ntxiv rau Buffer RL2 lossis Buffer RW2.
Kev pom zoo: Ua raws li cov lus qhia, ntxiv qhov tseeb ntim ntawm ethanol rau Buffer RL2 thiab Buffer RW2 thiab sib tov zoo ua ntej siv cov khoom siv.
6. Kev siv cov ntaub so ntswg tsis tsim nyog.
Kev pom zoo: Siv 10-20 mg ntawm cov ntaub so ntswg los yog (1-5) × 106hlwb ib 500 μl tsis RL1, raws li kev siv cov ntaub so ntswg ntau tuaj yeem ua rau txo qis RNA rho tawm.
7. Tsis tsim nyog elution ntim lossis tsis tiav elution.
Kev pom zoo: Lub elution ntim ntawm purification kem yog 50-200 μl;Yog tias cov nyhuv elution tsis txaus siab, nws raug nquahu kom txuas ntxiv chav tsev kub tso sijhawm tom qab ntxiv preheated RNase-Free ddH2O, piv txwv li rau 5-10 min.
8.Cov purification kem muaj ethanol residue tom qab Buffer RW2 ntxuav.
Kev pom zoo: Yog tias muaj ethanol residue tom qab Buffer RW2 ntxuav, khoob raj centrifugation rau 1min, lub sij hawm rau lub raj khoob centrifugation ua hauj lwm yuav nce mus rau 2min, los yog purification kem yuav muab tso rau hauv chav tsev kub rau 5 min kom txaus tshem tawm cov seem ethanol.
Purified RNA yog degraded
Qhov zoo ntawm RNA purified yog muaj feem xyuam rau tej yam xws li kev khaws cia ntawm tus qauv, RNase contamination, thiab manipulation, thiab lwm yam.
1. Cov qauv ntaub so ntswg tsis khaws cia rau lub sijhawm.
Kev pom zoo: Yog tias tsis siv cov ntaub so ntswg los yog cov hlwb tsis siv raws sij hawm tom qab sau, tam sim ntawd cryopreserve ntawm -80 ° C los yog kua nitrogen.Txhawm rau rho tawm RNA, siv cov ntaub so ntswg tshiab lossis cov qauv ntawm tes thaum twg los tau.
2. Rov qab khov-thawing ntawm cov ntaub so ntswg kuaj.
Kev pom zoo: Thaum khaws cov ntaub so ntswg, nws yog qhov zoo tshaj plaws los txiav lawv rau hauv me me rau kev khaws cia, thiab tshem tawm ib qho ntawm cov khoom thaum siv lawv kom tsis txhob rov khov-thawing ntawm cov qauv thiab degradation ntawm RNA.
3. RNase yog qhia los yog tsis hnav hnab looj tes, qhov ncauj qhov ntswg, thiab lwm yam thaum lub sijhawm ua haujlwm.
Kev pom zoo: RNA extraction thwmsim yog ua tau zoo tshaj plaws nyob rau hauv cais RNA manipulation chav thiab lub rooj yog cleared ua ntej qhov kev sim.
Hnav cov hnab looj tes thiab lub qhov ncauj qhov ntswg pov tseg thaum lub sijhawm sim ua kom txo qis RNA degradation los ntawm kev qhia txog RNase.
4. Cov tshuaj tiv thaiv kab mob muaj RNase thaum siv.
Kev Pom Zoo: Hloov nrog Cov Tsiaj Tag Nrho Tag Nrho RNA cais tawm tshiab rau cov kev sim cuam tshuam.
5. Cov raj centrifuge, cov lus qhia, thiab lwm yam siv hauv RNA manipulation yog paug nrog RNase.
Kev pom zoo: Paub meej tias cov raj centrifuge, cov lus qhia, pipettes, thiab lwm yam siv hauv RNA extraction yog txhua yam RNase-Free.
Purified tau txais RNA cuam tshuam cov kev sim hauv qab
RNA purified los ntawm purification kem, yog cov ntsev ions, cov ntsiab lus ntawm cov protein ntau dhau lawm yuav cuam tshuam rau cov kev sim qis qis, xws li: thim rov qab, Northern Blot li al.
1. Cov elutioned RNA muaj ntsev ion residues.
Kev pom zoo: Paub meej tias qhov tseeb ntim ntawm ethanol tau ntxiv rau Buffer RW2 thiab ua 2 purification kem ntxuav ntawm qhov ceev centrifugal qhia rau kev ua haujlwm;yog tias muaj cov ntsev ion residue, tawm hauv kab purification mus rau Buffer RW2 rau 5 min ntawm chav tsev kub thiab ua centrifugation kom loj tshaj qhov kev tshem tawm cov ntsev paug.
2. Ethanol residue nyob rau hauv elutioned RNA.
Kev pom zoo: Paub meej tias tom qab tsis RW2 ntxuav, ua lub raj khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob, ua rau lub qhov khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob, ua kom lub sij hawm ntawm lub raj khoob centrifugation mus rau 2 min.
