ua raws li daim ntawv cog lus ", ua raws li cov kev xav tau ntawm kev ua lag luam, koom nrog kev sib tw ua lag luam los ntawm nws cov khoom zoo ib yam li muab kev txhawb nqa ntau dua thiab zoo heev rau cov neeg siv khoom kom lawv ua tus yeej loj.Kev nrhiav ntawm lub tuam txhab, yog twv yuav raug hu cov neeg siv khoom txaus siab rau Tuam Tshoj Cov Khoom Tshiab Zoo Tshaj Muag Viral DNA & Rna Extraction Kit, Peb tau nthuav peb cov lag luam mus rau hauv lub teb chaws Yelemees, Qaib ntxhw, Canada, USA, Indonesia, Is Nrias teb, Nigeria, Brazil thiab lwm yam cheeb tsam ntawm lub ntiaj teb no.Peb tab tom ua haujlwm hnyav los ua ib tus neeg muab khoom zoo tshaj plaws thoob ntiaj teb.
ua raws li daim ntawv cog lus ", ua raws li cov kev xav tau ntawm kev ua lag luam, koom nrog kev sib tw ua lag luam los ntawm nws cov khoom zoo ib yam li muab kev txhawb nqa ntau dua thiab zoo heev rau cov neeg siv khoom kom lawv ua tus yeej loj.Kev nrhiav ntawm lub tuam txhab, yog twv yuav raug hu cov neeg siv khoom txaus siab rauTuam Tshoj Rna & DNA Extraction Kit thiab Nucleic Acid Extraction, Peb tab tom nrhiav rau pem hauv ntej los tsim kom muaj kev sib raug zoo nrog koj raws li peb cov khoom zoo thiab cov kev daws teeb meem, tus nqi tsim nyog thiab kev pabcuam zoo tshaj plaws.Peb cia siab tias peb cov khoom yuav ua rau koj muaj kev zoo siab thiab nqa kev zoo nkauj.
Phau ntawv qhia:Plant Total RNA Isolation Kit Plus Phau Ntawv Qhia

ua raws li daim ntawv cog lus ", ua raws li cov kev xav tau ntawm kev ua lag luam, koom nrog kev sib tw ua lag luam los ntawm nws cov khoom zoo ib yam li muab kev txhawb nqa ntau dua thiab zoo heev rau cov neeg siv khoom kom lawv ua tus yeej loj.Kev nrhiav ntawm lub tuam txhab, yog twv yuav raug hu cov neeg siv khoom txaus siab rau Tuam Tshoj Cov Khoom Tshiab Zoo Tshaj Muag Viral DNA & Rna Extraction Kit, Peb tau nthuav peb cov lag luam mus rau hauv lub teb chaws Yelemees, Qaib ntxhw, Canada, USA, Indonesia, Is Nrias teb, Nigeria, Brazil thiab lwm yam cheeb tsam ntawm lub ntiaj teb no.Peb tab tom ua haujlwm hnyav los ua ib tus neeg muab khoom zoo tshaj plaws thoob ntiaj teb.
Tuam Tshoj Cov Khoom Tshiab Tuam Tshoj Rna & DNA Extraction Cov Khoom Siv thiab Nucleic Acid Extraction, Peb tab tom nrhiav rau pem hauv ntej los tsim kom muaj kev sib raug zoo nrog koj raws li peb cov khoom zoo thiab cov kev daws teeb meem, tus nqi tsim nyog thiab cov kev pabcuam zoo tshaj plaws.Peb cia siab tias peb cov khoom yuav ua rau koj muaj kev zoo siab thiab nqa kev zoo nkauj.

Lub kem ntsaws

Tom qab kab txuas, RNA tawm los yog txo qis los yog tsis tuaj yeem ua kom huv RNA, thiab qhov tau txais RNA pawg tsawg.

Common ua Analysis:

1. Cov qauv tawg tsis zoo.

Cov qauv tawg tsis tas ua rau DNA-CLEANING COLUMN raug thaiv, thaum cuam tshuam rau RNA yield thiab zoo.Peb pom zoo kom ceev kev sib tsoo ua haujlwm hauv cov kua nitrogen txaus thaum koj tsoo cov qauv, Sim zom cov qauv ntawm phab ntsa, cell membrane thiab lwm yam ntaub so ntswg.Rau cov qauv cog ntawm polyol polysaccharides, peb xav kom koj siv Plant Total RNA ISOLATION KIT PLUS.

2. Thaum nqus cov qauv sib cais supernatant nrog DNA-Tshuaj Kem, cov cell fragmented precipitate tuaj yeem nqus tau.

Lub cell fragmented sediments coj mus yuav ua rau RNA-Tsuas Kem uas yuav raug thaiv thaum ua haujlwm RNA adsorption (saib kauj ruam 6).Peb pom zoo kom koj ua tib zoo thaum nqus cov supernatant no kom tsis txhob muaj cov khib nyiab ntawm tes raug nqus.

3. Tus nqi pib thawj yog ntau dhau.

Kev siv cov qauv ntau dhau yuav ua rau tsis tiav cov qauv tawg lossis tsis tiav cell lysis los ntawm Buffer PSL1, uas ua rau muaj kev cuam tshuam ntawm purification kem thaum purification.Plant Total RNA Isolation Kit Txhua tus qauv ua haujlwm huv si yog 50 mg.Rau cov qauv cog ntawm polyol polysaccharides, peb xav kom koj sim Plant Total RNA ISOLATION KIT PLUS.

4. Qhov kub ntawm lub centrifuge yog qis heev.

Tag nrho RNA kev rho tawm thiab kev ua kom huv yog nqa tawm ntawm chav tsev kub (20-25°C), tshwj tsis yog tias cov qauv ntaub so ntswg tawg los ntawm cov kua nitrogen.Qhov kub ntawm qee qhov cryogenic centrifuges qis dua 20℃, uas yuav ua rau muaj kev thaiv ntawm DNA-Tshuaj Kem thiab / lossis RNA-Tsuas Kab.Yog hais tias qhov no tshwm sim, teem lub centrifuge kub rau 20-25℃, thiabxyuas kom cov lysis sib tov thiab/los yog cov ethanol-ntxiv supernatant tau preheated rau 37°C.

Tsis muaj RNA rho tawm lossis RNA tawm los tsawg

Feem ntau muaj ntau yam uas cuam tshuam rau kev rov ua haujlwm zoo, xws li: qauv RNA cov ntsiab lus, txoj kev ua haujlwm, qhov ntim elution, thiab lwm yam.

Kev tshuaj xyuas ntawm cov laj thawj uas tshwm sim hauv qab no:

1.Ib da dej dej khov lossis kub qis (4 ° C) centrifugation tau ua thaum lub sijhawm ua haujlwm.

Tswv yim: Ua haujlwm ntawm chav tsev kub (15-25°C) nyob rau hauv tag nrho cov txheej txheem, tsis txhob ua dej khov da dej thiab tsis kub centrifugation.

2.Lub RNA tau raug degraded vim yog tsis tsim nyog preservation ntawm cov qauv los yog mus sij hawm ntev khaws cia ntawm tus qauv.

Cov lus pom zoo: Cov qauv sau tshiab yuav tsum tau khov sai sai hauv cov kua nitrogen, thiab tom qab ntawd khaws cia ntawm -80 ° C rau lub sijhawm ntev, tsis txhob rov khov thiab thawing ntawm cov qauv;lossis tam sim ntawd tsau cov qauv hauv RNA stabilizer RNAlater tov (cov qauv tsiaj txhu).

3.Cov qauv tsis txaus fragmentation thiab lysis ua rau blockage ntawm purification kem.

Tswv yim: Thaum sib tsoo cov ntaub so ntswg, thov xyuas kom meej tias cov ntaub so ntswg yog av txaus, thiab sai sai hloov mus rau lub pre-npaj Buffer PSL1 (kom paub meej tias qhov tseeb feem ntawm β-ME tau ntxiv, saib cov kauj ruam 1 ntawm cov txheej txheem).

4.Qhov eluent tau ntxiv tsis raug.

Tswv yim: Xyuas kom tseeb tias RNase-Free ddH2O yog dripped rau hauv nruab nrab ntawm cov purification kem membrane.

5.Qhov tseeb ntim ntawm ethanol tsis tau ntxiv rau Buffer PSL2 lossis Buffer PRW2.

Tswv yim: Thov ua raws li cov lus qhia, ntxiv qhov tseeb ntim ntawm ethanol rau Buffer PSL2 thiab Buffer PRW2 thiab sib tov zoo ua ntej cov khoom siv.

6.Cov qauv ntawm cov ntaub so ntswg tsis tsim nyog.

Tswv yim: Siv 50 mg ntawm cov ntaub so ntswg rau 500 μl ntawm Buffer PSL1.Kev siv cov ntaub so ntswg ntau dhau yuav txo tau cov RNA rho tawm thiab qhov purity ntawm qhov tshwm sim RNA kuj yuav raug txo.Peb xav kom pom zoo tias thawj zaug koob tshuaj yuav tsum tsis txhob ntau tshaj 50 mg rau RNA rho tawm haujlwm.

7.Inappropriate elution ntim lossis tsis tiav elution.

Tswv yim: Lub ntim eluent ntawm purification kem yog 50-200 μl;Yog tias cov nyhuv elution tsis txaus siab, nws raug nquahu kom ncua lub sijhawm ntawm chav tsev kub tom qab ntxiv preheated RNase-Free ddH2O, xws li 5-10min.

8.Cov purification kem muaj ethanol residue tom qab ntxuav nrog BufferPRW2.

Cov lus qhia: Yog tias lub raj khoob yog centrifuged rau 1 min thiab tseem muaj ethanol seem tom qab ntxuav hauv Buffer PRW2, koj tuaj yeem nce lub sij hawm ntawm lub raj khoob centrifugation mus rau 2 min, los yog tso cov purification kem ntawm chav tsev kub rau 5 min kom tshem tawm cov ethanol seem.

9.Cov khoom siv tau siv tsis raug.

Tswv yim: Rau cov qauv cog ntawm polyphenolic polysaccharides, siv cov khoom siv xws li Plant Total RNA Isolation Kit yuav tsis tuaj yeem tau txais cov qauv RNA zoo tagnrho.Peb pom zoo kom koj siv Plant Total RNA IsolationKit Plus, uas yog tsim tshwj xeeb rau cov qauv cog polyphenolic polysaccharide.Cov khoom siv tshwj xeeb tsim los rau rho tawm RNA los ntawm polyphenol thiab polysaccharide cog cov qauv.

OD260 / OD280 tus nqi qis

RNA elution nrog ddH2O thiab siv rau kev nyeem ntawv spectrophotometer ua rau qis OD260 / OD280 qhov tseem ceeb.Peb pom zoo kom siv 10 mM Tris-HCl, pH 7.5 (tshwj tsis yog RNase-Free ddH2O rau elute RNA) kom tau txais cov txiaj ntsig zoo OD260/OD280, saib "RNA Concentration thiab Purification Assays" ntawm nplooj 19.

Lub purified RNA yog degraded

Qhov zoo ntawm purified RNA muaj feem xyuam rau yam xws li cov qauv khaws cia, RNase contamination, thiab manipulation.

Kev tshuaj xyuas ntawm cov laj thawj uas tshwm sim:

1.Cov ntaub so ntswg tsis tau muab khaws cia rau hauv lub sijhawm tom qab sau.

Kev pom zoo: Yog tias cov ntaub so ntswg tsis siv rau lub sijhawm tom qab sau, thov khaws cia rau hauv cov kua nitrogen ntawm qhov kub thiab txias tam sim ntawd lossis hloov mus rau -80 ° C rau lub sijhawm ntev cia tom qab ceev ceev hauv cov kua nitrogen, lossis tam sim ntawd tso cov qauv hauv RNA stabilizer RNAlater tov (cov qauv tsiaj txhu).Rau RNA rho tawm, sim siv cov ntaub so ntswg tshiab tshiab.

2.Repeated khov thiab thawing ntawm cov ntaub so ntswg kuaj.

Cov lus qhia: Thaum khaws cov ntaub so ntswg, nws yog qhov zoo tshaj plaws los txiav rau hauv me me rau kev khaws cia, thiab tshem tawm ib feem ntawm lawv thaum siv lawv kom tsis txhob degradation ntawm RNA los ntawm kev rov ua kom khov thiab thawing ntawm cov qauv.

3.RNase tau qhia hauv chav ua haujlwm lossis tsis hnav cov hnab looj tes, qhov ncauj qhov ntswg, thiab lwm yam.

Tswv yim: RNA extraction thwmsim yog ua tau zoo tshaj plaws nyob rau hauv cais RNA kev khiav hauj lwm, thiab lub chav kuaj lub rooj yuav tsum tau ntxuav ua ntej qhov kev sim, thiab pov tseg hnab looj tes thiab lub qhov ncauj qhov ntswg yuav tsum tau hnav thaum lub sij hawm sim kom tsis txhob RNA degradation los ntawm kev qhia RNase mus rau qhov loj tshaj plaws.

4.Cov reagent muaj paug los ntawm RNase thaum siv.

Tswv yim: Hloov nrog ib tug tshiab series ntawm cov nroj tsuag tag nrho RNA extraction kits rau lwm yam thwmsim.

5.Cov centrifuge raj thiab pipette cov lus qhia siv rau RNA manipulation yog kis nrog RNase.

Cov lus qhia: Xyuas kom tseeb tias cov raj centrifuge, cov lus qhia pipette, pipettes, thiab lwm yam siv hauv RNA extraction yog txhua yam RNase-Free.

Phau ntawv qhia:

Plant Total RNA Isolation Kit Plus Phau Ntawv Qhia