ⅠNucleic acidikev taw qhia
Nucleic acid tau muab faib ua deoxyribonucleic acid (DNA) thiab ribonucleic acid (RNA), uas RNA muab faib ua ribosomal RNA (rRNA), tub txib RNA (mRNA) thiab hloov RNA (tRNA) raws li nws cov haujlwm sib txawv.DNA feem ntau yog concentrated nyob rau hauv lub nucleus, mitochondria thiab chloroform, thaum RNA feem ntau yog faib nyob rau hauv lub cytoplasm.Raws li cov ntaub ntawv hauv paus ntawm noob caj noob ces, nucleic acid extraction plays lub luag haujlwm tseem ceeb hauv kev tshawb fawb molecular biology thiab kev kuaj mob molecular.Qhov concentration thiab purity ntawm nucleic acid extraction yuav ncaj qha cuam tshuam rau PCR tom ntej, sequencing, vector tsim, enzyme digestion thiab lwm yam kev sim.
Ⅱ Nucleic acid extraction thiab purification txoj kev
① Phenol/chloroform extraction method
Phenol / chloroform extraction yog ib txoj kev classical rau DNA extraction, uas feem ntau yog siv ob hom organic solvents los kho cov qauv, dissolving DNA raws nucleic acid nyob rau hauv dej theem, lipids nyob rau hauv lub organic theem, thiab cov proteins ntawm ob theem.Txoj kev no muaj qhov zoo ntawm tus nqi qis, siab purity thiab cov nyhuv zoo.Qhov tsis zoo yog kev ua haujlwm nyuaj thiab lub sijhawm ntev.
② Trizol txoj kev
Trizol txoj kev yog ib txoj kev classical rau RNA rho tawm.Txoj kev Trizol muab faib ua aqueous theem thiab organic theem tom qab centrifugation nrog chloroform, nyob rau hauv uas RNA yog yaj nyob rau hauv lub aqueous theem, lub aqueous theem yog pauv mus rau ib tug tshiab EP raj, nag lossis daus yog tau tom qab ntxiv isopropanol, thiab ces ethanol purification.Txoj kev no yog tsim rau RNA rho tawm los ntawm cov ntaub so ntswg tsiaj, hlwb thiab kab mob.
③ Centrifugal kem purification txoj kev
Centrifuge kem purification txoj kev tuaj yeem adsorb DNA tshwj xeeb los ntawm cov khoom siv tshwj xeeb silicon matrix adsorption, thaum RNA thiab cov protein tuaj yeem hla tau zoo, thiab tom qab ntawd siv cov ntsev qis PH los ua ke cov nucleic acid, cov ntsev tsawg PH tus nqi elution cais thiab purify nucleic acid.Qhov zoo yog siab purification concentration, siab stability, tsis xav tau cov organic hnyav, thiab tus nqi qis.Qhov tsis zoo yog tias nws yuav tsum tau centrifuged kauj ruam ntawm kauj ruam, ntau cov kauj ruam ua haujlwm.
④ Magnetic hlaws txoj kev
Txoj kev sib nqus hlaws yog cais cov qauv ntawm tes los ntawm lysate, tso cov nucleic acid hauv cov qauv, thiab tom qab ntawd cov nucleic acid molecules tshwj xeeb yog adsorbed rau saum npoo ntawm cov hlaws hlau nplaum, thaum impurities xws li cov proteins thiab suab thaj raug tso rau hauv. cov kua.Los ntawm cov kauj ruam ntawm cov ntaub so ntswg splitting, sib nqus hlaws khi nrog nucleic acid, nucleic acid ntxuav, nucleic acid elution, thiab lwm yam, cov ntshiab nucleic acid yog thaum kawg.Qhov zoo yog kev ua haujlwm yooj yim thiab siv sijhawm luv luv, tsis tas yuav tsum tau ua cov kauj ruam centrifugation.Nws muaj cov kev cai qis thiab tuaj yeem paub txog kev ua haujlwm tsis siv neeg thiab loj.Kev sib xyaw tshwj xeeb ntawm cov hlaws sib nqus thiab cov nucleic acid ua rau cov kua qaub uas tau muab rho tawm nrog siab concentration thiab purity.Qhov tsis zoo yog tias tus nqi lag luam tam sim no kuj kim heev.
⑤ Lwm txoj hauv kev
Ntxiv nrog rau plaub txoj kev saum toj no, muaj cov boiling cracking, concentrated ntsev txoj kev, anionic detergent txoj kev, ultrasonic txoj kev thiab enzymatic txoj kev, thiab lwm yam.
Ⅲ Hom ntawm nucleic acid extraction
Foregene muaj lub ntiaj teb ua Direct PCR platform, ob-kem RNA Isolation platform (DNA nkaus xwb + RNA nkaus xwb).Cov khoom tseem ceeb suav nrog DNA / RNA cov khoom siv cais tawm, PCR thiab Direct PCR reagents molecular lab reagents series.
① Tag nrho RNA rho tawm
Tag nrho RNA rho tawm cov qauv muaj xws li cov ntshav, hlwb, cov ntaub so ntswg tsiaj, nroj tsuag, kab mob, thiab lwm yam. High purity thiab siab concentration ntawm tag nrho RNA yuav tau los ntawm tag nrho cov RNA extraction, uas yuav siv tau nyob rau hauv RT-PCR, nti tsom xam, in vitro translation, molecular cloning, Dot Blot thiab lwm yam kev sim.
Foregene cuam tshuamRNA Isolation Cov Khoom Siv
Tsiaj Tag Nrho RNA Isolation Kit--Ceev ceev thiab ua tau zoo extract high-purity thiab high-zoo tag nrho RNA los ntawm ntau yam tsiaj cov ntaub so ntswg.
Cell Total RNA Isolation Kit--Ua kom huv thiab zoo tag nrho RNA tuaj yeem tau txais los ntawm ntau lub hlwb hauv 11 feeb.
Plant Total RNA Isolation Kit--Ceev nrooj tshem tawm tag nrho cov RNA zoo los ntawm cov qauv cog nrog cov ntsiab lus tsis tshua muaj polysaccharide thiab polyphenol.
Viral RNA Isolation Kit--Kev cais tawm sai sai thiab ntxuav cov kab mob RNA los ntawm cov qauv xws li plasma, ntshav, cell-free lub cev kua thiab cell culture supernatants.
② Genomic DNA rho tawm
Genomic DNA extraction qauv muaj xws li av, quav, ntshav, hlwb, tsiaj cov ntaub so ntswg, nroj tsuag, kab mob, thiab lwm yam. Genomic DNA extraction yuav siv tau nyob rau hauv enzyme digestion, DNA tsev qiv ntawv tsim, PCR, antibody npaj, Western blot hybridization tsom xam, noob nti, siab. -Txoj kev sib sau ua ke thiab lwm yam kev sim.
Foregene cuam tshuamDNA Isolation Kits
Animal Tissue DNA Isolation Kit--Kev rho tawm sai thiab ua kom huv ntawm genomic DNA los ntawm ntau qhov chaw, xws li tsiaj cov ntaub so ntswg, hlwb, thiab lwm yam.
Ntshav DNA Midi Kit (1-5ml)- Ceev ceev purify zoo genomic DNA los ntawm anticoagulated ntshav (1-5ml).
Buccal Swab/FTA Card DNA Isolation Kit- Ceev ceev purify high-zoo genomic DNA los ntawm buccal swab / FTA Card kuaj.
Cog DNA Isolation Kit- Ua kom huv sai sai thiab tau txais txiaj ntsig zoo genomic DNA los ntawm cov qauv cog (xws li polysaccharides thiab polyphenol cog qauv)
③ Plasmid extraction
Plasmid yog ib hom kab me me molecule DNA nyob rau hauv cov hlwb, uas yog ib qho kev thauj mus rau DNA recombination.Txoj kev ntawm plasmid extraction yog tshem tawm RNA, cais plasmid los ntawm cov kab mob genomic DNA, thiab tshem tawm cov protein thiab lwm yam impurities kom tau txais cov plasmid ntshiab.
General Plasmid Mini Cov Khoom Siv- Ceev ceev purify high-zoo plasmid DNA los ntawm cov kab mob hloov mus rau niaj hnub molecular biology thwmsim xws li transformation thiab enzyme digestion
④ Lwm hom kev rho tawm, miRNA rho tawm, thiab lwm yam.
Tsiaj MiRNA Isolation Kit- Ceev ceev thiab fficiently rho me me RNA fragments ntawm 20-200nt miRNA, siRNA, snRNA los ntawm ntau yam tsiaj cov ntaub so ntswg thiab hlwb
Ⅳ Kev xav tau rau nucleic acid extraction thiab purification tshwm sims
① Los xyuas kom meej qhov kev ncaj ncees ntawm thawj cov qauv ntawm nucleic acid.
② Txo qhov cuam tshuam ntawm cov protein, suab thaj, lipids thiab lwm yam macromolecules
③ Yuav tsum tsis muaj cov kuab tshuaj organic lossis siab concentration ntawm cov hlau ions uas tuaj yeem cuam tshuam cov enzyme hauv cov qauv nucleic acid.
④ RNA thiab lwm yam kab mob nucleic acid yuav tsum raug tshem tawm thaum rho tawm DNA, thiab rov ua dua.
Post lub sij hawm: Nov-24-2022
