Lub hauv paus rau phau ntawv
● Qhov tseem ceeb ntawm Genomic DNA Extraction
Nyob rau hauv lub realm ntawm molecular biology, extraction ntawm genomic DNA yog qhov ua rau cov ntawv thov, los ntawm kev tshawb fawb txog kev kuaj mob thiab tshuaj kho tus kheej. Cov txheej txheem DNOOM siv cuam tshuam nrog DNACHED DNA los ntawm cov kab mob caj ces, muab cov txuj ci tseem ceeb, thiab kev txhim kho biotechnological. Nrog rau kev ntseeg siab thov kom tshawb fawb Genomic, lub luag haujlwm ntawm kev ntseeg siabGenomic DNA Extraction Cov Khoom Sivyeej tsis tau muaj ntau ntxiv pivotal. Cov khoom siv no rub tawm cov txheej txheem rho tawm, kom muaj txiaj ntsig zoo thiab kev ua kom dawb huv ntawm DNA, qhov tseem ceeb rau cov ntawv thov hauv qab.
Cov txheej txheem sau ua qauv
Cov qauv ntawm cov qauv rau DNA Extraction
Kev ncaj ncees ntawm Genomic DNA pib nrog kev sau cov qauv siab - cov qauv zoo. Cov chaw sib xws suav nrog cov ntshav, ntaub so ntswg, qaub ncaug, thiab cov pob zeb ci ci. Txhua tus qauv hom muaj cov txiaj ntsig tshwj xeeb; Piv txwv li cov ntshav cov ntshav yog nplua nuj nyob hauv DNA tab sis yuav tsum tau ua tib zoo saib, thaum qaub ncaug thiab buccal so - kev xaiv. Xaiv cov qauv uas tsim nyog yog qhov tseem ceeb, tshwj xeeb tshaj yog thaum siv cov khoom siv ntawm tes ntawm tes, uas yuav yog qhov zoo rau cov xovtooj ntawm cov xovtooj no, kom muaj txiaj ntsig ntau tshaj plaws thiab kev coj dawb huv.
● Cov kev coj ua zoo tshaj plaws rau kev coj ua piv txwv
Ua kom ntseeg tau cov qauv kev ncaj ncees yog qhov tseem ceeb rau kev ua tiav DNA extraction. Ib puag ncig yuav tsum muab cov hnoos qeev no yuav tsum tau muab cov tshuaj sib xyaw - ib puag ncig dawb, khaws cia rau ntawm qhov ntsuas kub uas tsim nyog, thiab daws nrog kev saib xyuas kom tiv thaiv kev ua kom tsis txhob muaj kev txwv. Cov kauj ruam no yog qhov tseem ceeb seb koj puas tau ua nrog cov hnoos qeev tshiab lossis cov kev tsis txaus siab tuaj yeem cuam tshuam txog qhov kawg DNA tawm thiab ua tau zoo, cuam tshuam rau kev sim ua ntej.
Cell lysis txoj kev
● Tshuaj thiab cov txuj ci dag cov tswv yim
Cov kauj ruam ntawm tes yog tseem ceeb hauv kev tso tawm cov qauv ntawm cov qauv cellular. Tshuaj lysis, koom tes nrog xab npum thiab cov enzymes, maj mam rhuav cov xov tooj ntawm tes thiab cov protein. Kev siv lub cev - xws li kev cuam tshuam txhua yam kev cuam tshuam thiab sonication-muaj kev siv zog ntau dua thiab tuaj yeem siv rau cov qauv ntsuas nruj. Kev xaiv ntawm txoj kev feem ntau yog nyob ntawm hom qauv thiab cov kev xav tau ntawm cov khoom siv DNO Genomic DNA. Cov khoom siv tsim cov khoom siv tsim kho kom ua kom tau cov khoom ua kom zoo, ntsuas qhov yooj yim thiab ua haujlwm tau zoo.
● Qhov tseem ceeb ntawm cov lysis hauv DNA tso tawm
Cov lus dag ua kom zoo dua tias cov khoom siv cell yog tawg, ua kom tiav qhov kev tso tawm ntawm kev daws teeb meem. Kaw cov lus ua haujlwm kom zoo ntawm cov DNA thiab paug los ntawm cov protein thiab cov lipids, uas yog qhov tshwj xeeb tshaj plaws hauv kev kho mob cell, qhov twg genomic kev ncaj ncees.
Tshem tawm cov protein thiab impurities
● Lub luag haujlwm ntawm cov kuab tshuaj hauv kev huv
Xa - lysis, tshem tawm cov protein thiab lwm yam tsis txaus ntseeg yog qhov tseem ceeb. Cov kuab tshuaj zoo li phenol - chloroform yog kev lig kev cai ua haujlwm kom tsis muaj txiaj ntsig thiab cais cov protein ntawm nucleic acids. Niaj hnub nimno Genomic DNA extraction cov khoom siv feem ntau yog siv silica - raws li cov mem tes lossis cov hlau sib nqus, uas khi cov tshuaj tiv thaiv thaum tso cai kom ntxuav tawm. Cov kauj ruam no yog qhov tseem ceeb heev hauv kev tau txais DNA ntshiab rau cov ntawv thov hauv qab thiab rhiab dhau.
● Centrifugation piav qhia
Centrifugation yog ib qib zuj zus los cais DNA los ntawm impurities post - lysis. Los ntawm kev thov centrifugal quab yuam, hnyav cellular cov khib nyiab thiab cov protein yog pelleted, thaum lub DNA tseem nyob hauv supernatant. Cov kauj ruam no feem ntau rov ua dua nyob rau hauv Tandem nrog cov kauj ruam lim kom txhim kho cov kev coj dawb huv thiab concentration ntawm extracted DNA.
DNA nag los cov txheej txheem
● Kev siv isopropanol thiab ethanol
Nag lossis daus yog cov txheej txheem classic rau kev rov qab los ntawm kev daws, siv nyob rau hauv ze li tag nrho cov genomic DNA cov khoom siv extraction. Cawv xws li isopropanol thiab ethanol pab kom DNA ua daus los ntawm kev txo qis nws solubility. Ib zaug precipitated, DNA cov ntawv pom kev clumps lossis filaments, muab kev pom cue ntawm kev ua tiav.
Vestizing DNA filaments
Lub xub ntiag ntawm DNA filaments lees paub siv nag ntsis thiab ua haujlwm ua qhov tseeb ntawm kev ua tiav. Cov kauj ruam no, txawm tias qhov yooj yim zoo li, yog qhov tseem ceeb rau kev ntseeg siab rau cov txheej txheem rho tawm, kom muaj DNA txaus lawm.
Ntxuav Daus Da Dna
Kuj tseem ceeb ntawm ethanol ntxuav
Ntxuav DNA pellets nrog ethanol ua haujlwm kom tshem tawm cov ntsev uas cov neeg muaj sia thiab tsis zoo. Cov kauj ruam no, txawm hais tias kev coj ua, yog qhov tseem ceeb hauv kev tiv thaiv kev ua paug uas tuaj yeem cuam tshuam cov txheej txheem downstream. Ethanol ntxuav yog cov txawj ntse tsis muaj cai nyob rau hauv txhua phau ntawv siv cov phau ntawv DNA los ntawm Genomic DNA Extraction cov txiaj ntsig los xyuas kom cov neeg siv tau ua tiav.
● Saib xyuas kev coj dawb huv ntawm DNA
Purity yog ib qho tseem ceeb raws li tawm los hauv DNA rho tawm. Residual impurities tuaj yeem txwv tsis tau enzymatic cov tshuaj tiv thaiv hauv cov ntawv thov nqes hav xws li PCR thiab ua ntu zus. Genomic DNA Extraction cov khoom siv hu ua kev coj dawb huv, muab cov kev cai thiab cov ntaub ntawv uas ua kom muaj kuab paug yog kom huv si.
Dissolving thiab npaj DNA
Xaiv Txoj Cai Zoo
Thaum cov DNA yog ntxuav, nws yog yaj nyob rau hauv ib qho kev haum tsis zoo, feem ntau yog cov dej tsis zoo lossis ua kom tsis huv. Qhov kev xaiv ntawm Buffer tuaj yeem cuam tshuam rau kev ruaj ntseg thiab kev ncaj ncees ntawm DNA, cuam tshuam nws ntev ntev - TEWANCE siv tau. Genomic DNA Extraction cov khoom siv feem ntau muab cov buffers ua kom haum rau cov ntawv thov hauv qis.
● Npaj DNA rau kev sim
Kev npaj kom tsim nyog ntawm DNA yog qhov tseem ceeb rau kev sim ua tiav, ua kom muaj kev sib raug zoo nrog cov txheej txheem qpr, thiab txuas ntxiv mus. Cov kev npaj zoo tshaj plaws ua rau muaj kev ncaj ncees DNA thiab kev ua tau zoo hauv cov ntawv thov no.
Kev Tswj Xyuas Zoo Ntawm Extracted DNA
● Cov tswv yim los ntsuas DNA zoo
Tom qab rho tawm, ntsuas DNA zoo yog qhov tseem ceeb. Spectrophotometry yog cov qauv nrov, ntsuas kev nqus ntawm 260 nm thiab muab cov ntaub ntawv hais txog DNA kev xav thiab huv. Ib qho ntxiv, Gel Electrophoresis tso cai rau kev pom ntawm DNA kev ncaj ncees, kuaj pom txhua qhov kev tsis txaus siab.
● Kawm txog kev nyeem ntawv Spectrophotometer nyeem
Spectrophotometer nyeem muab kev pom rau hauv kev siab thiab huv ntawm DNA. Cov 260/280 piv ze rau 1.8 qhia txog DNA ntshiab, thaum kev sib txawv qhia kom pom tseeb. Cov kev nyeem no yog qhov tsis zoo nyob rau hauv kom ntseeg tau tias cov DNA yog haum rau cov ntawv thov rhiab.
Cov kev cov nyom hauv DNA rho tawm
Cov teeb meem thiab kev daws teeb meem
Txawm hais tias nce qib, cov nyom hauv DNA rho tawm tsis kam, suav nrog cov txiaj ntsig qis, thiab kev tsis sib thooj. Nkag siab txog cov teeb meem no thiab ua raws cov txheej txheem daws teeb meem-feem ntau muab los ntawm cov tuam ntxhab-yog qhov tseem ceeb rau kev ua tiav.
● Txawv sib txawv nrog cov qauv sib txawv
Cov piv txwv sib txawv hom kev sib tw cov kev sib tw tshwj xeeb hauv DNA extraction, xws li nce qib DNA cov ntsiab lus thiab muaj nyob ntawm Insibitors. Genomic DNA Extraction Cov Khoom Siv Tsim Qauv Tsim Kho cov khoom siv los daws cov kev hloov pauv no, muab cov kev daws teeb meem rau ntau yam.
Kev nce qib hauv DNA rho thev naus laus zis
Kev Tsim Kho Hauv Cov Khoom Siv Tawm
Kev nce qib tas mus li hauv Genomic DNA extraction cov khoom tau tau yooj yim txheej txheem, txhim kho kev ua haujlwm, nrawm, thiab kev ntseeg tau. Cov kev hloov tshiab no suav nrog kev siv tshuab ua, kev sib nqus tshuab, thiab kev sib xyaw ua ke rau cov kev xav tau siab ntawm kev tshawb fawb garomic.
At Yav Tom Ntej Cov Qauv Hauv Genomic Sciences
Raws li Scomic Sciences ua ntej, qhov kev thov rau siab ntev - Cov tshuaj tiv thaiv DNA zoo DNA yuav loj tuaj. Cov ncauj lus yav tom ntej muaj ntau cov txheej txheem ntau dua, kev siv tshuab, thiab kev sib xyaw ua ke nrog cov kev ntsuas kom muaj txiaj ntsig zoo thiab muaj txiaj ntsig zoo nkauj.
BlueKit: Pioneer nyob rau hauv cov kev kho mob ntawm cev thiab cellular
Jiangsu Hadgene, hauv qab homCov bluekit, sawv ntsug ntawm qhov ua ntej ntawm kev kho mob ntawm kev txhim kho thiab kev kho mob cell. Lub taub hau nyob hauv Suzhou nrog cov chaw ntxiv nyob rau hauv Shenzhen thiab Shanghai, thiab lub vev xaib tom ntej no hauv North Carolina, toj siab txuas ntxiv nws mus txog thoob ntiaj teb. Cov blukit®Cov khoom muaj cov kev daws teeb meem rau kev tswj hwm ntawm tes, txhawb txoj kev txhim kho ntawm cov kev mob cell siab zoo li lub tsheb - T thiab TCR - t. Dhau los ntawm cov mob siab rau lub platform muaj kua qaub thiab QC xeem, BlueKit yog hloov kho ntawm kev kho mob cellular, nqa cov khoom lag luam xa khoom sai dua thiab tau zoo dua.
Lub Sijhawm Post: 2024 - 12 - 05 15:07:02
