Qhov sib txawv tseem ceeb ntawm DNA thiab RNA kev rho tawm yog qhov kev rho tawm DNA yog txheej txheem uas ib hom nucleic acid hu ua DNA tau muab rho tawm los ntawm cov cell lom, thaum RNA cais tawm yog txheej txheem uas hom nucleic acid hu ua RNA yog muab rho tawm los ntawm ib tug kab mob lom.
Nucleic acid cais tawm yog qhov pib ntawm qhov chaw loj ntawm cov ntawv sau cia. Qhov zoo ntawm nucleic acids nyob rau hauv cov qauv pib yog qhov tseem ceeb rau kev ua tiav ntawm cov ntawv thov hauv qab no. Nucleic acid cais tawm tuaj yeem txhais tau tias yog cov kauj ruam kom tau txais nucleic acid hauv cov qauv purity uas tuaj yeem siv rau cov kauj ruam sib txawv hauv qab. Lub hom phiaj ntawm nucleic acid extraction yog kom disintegrate cell membrane thiab ua tiav qhov siab tshaj plaws tshem tawm lipids thiab proteins kom tau cov ntshiab nucleic acids xws li DNA los yog RNA.
YDNA Isolation yog dab tsi?
DNA rho tawm yog ib txoj kev niaj hnub uas ib hom nucleic acid hu ua DNA yog muab rho tawm los ntawm ib tug lom cell. Kev cais DNA yog ua los ntawm kev rho tawm DNA los ntawm cov ntshav, cov ntaub so ntswg khov, thiab cov ntaub so ntswg paraffin blocks. Muaj peb theem tseem ceeb hauv kev cais DNA, suav nrog cell lysis, rho DNA, thiab nag lossis daus. Hauv cell lysis, cov hlwb hauv cov qauv sib cais los ntawm ib leeg feem ntau los ntawm kev siv lub cev xws li kev sib tsoo lossis vortexing thiab muab tso rau hauv cov tshuaj uas muaj ntsev. Feem ntau, cov txiaj ntsig zoo ntawm sodium ions hauv cov ntsev pab tiv thaiv cov pab pawg tsis zoo phosphate uas khiav raws cov caj qaum ntawm DNA. Tom qab ntawd, ib qho tshuaj ntxuav tau ntxiv los rhuav tshem cov lipids hauv cell membrane thiab nuclei, uas tso tawm DNA vim tias cov membranes no cuam tshuam.
Daim duab 01: DNA cais
DNA rho tawm yog ua kom tau txais cov qauv DNA huv. Feem ntau, ib qho protease enzyme ntxiv rau degrade DNA-txuas nrog cov proteins thiab lwm cov cellular proteins. Ntxiv mus, qee qhov ntawm cov khib nyiab ntawm tes tuaj yeem raug tshem tawm los ntawm kev lim cov qauv. Thaum kawg, DNA yog precipitated los ntawm kev siv cawv. Tsis tas li ntawd, yog tias muaj ntau ntawm DNA, ib qho stingy dawb precipitate thaum kawg ntawm txoj kev tuaj yeem pom.
RNA Isolation yog dab tsi?
RNA rho tawm yog ib txoj kev uas ib hom nucleic acid hu ua RNA yog muab rho tawm los ntawm ib tug lom cell. Cov txheej txheem no feem ntau nyuaj los ntawm qhov muaj ribonuclease enzymes hauv cov hlwb thiab cov ntaub so ntswg uas degrade RNA. Ntau txoj hauv kev siv hauv molecular biology los cais RNA los ntawm cov qauv, xws li guanidinium-thiocyanate-phenol-chloroform extraction.
Daim duab 02: RNA Isolation
RNA kev rho tawm feem ntau muaj ob peb kauj ruam, suav nrog cell lysis thiab homogenization, quenching ntawm biochemical txheej txheem, nucleic acid muab faib, RNA retrieval, crude purification, thiab ntsuam xyuas qhov zoo ntawm RNA. Cov ntawv lim ntawv-raws li lysis thiab elution txoj kev muaj peev xwm dhau los ntawm RNA rho tawm. Tsis tas li ntawd, RNA rho tawm hauv cov kua nitrogen los ntawm kev siv tshuaj khib thiab pestle kuj yog ib txoj hauv kev zoo heev vim nws tiv thaiv kev ua haujlwm ribonuclease.
Dab tsi zoo sib xws ntawm DNA thiab RNA cais tawm?
- DNA thiab RNA cais tawm yog ob hom nucleic acid cais cov txheej txheem.
- Ob txoj kev muaj cov kauj ruam yooj yim zoo sib xws, xws li cell lysis, extraction, thiab purification.
- Cov txheej txheem no cais cov nucleic acid tseem ceeb uas muaj ntau yam kev siv qis qis.
- Lawv tau ua hauv kev teeb tsa hauv chav kuaj.
- Lawv tuaj yeem ua tau siv cov khoom siv tsim khoom.
Qhov txawv ntawm DNA thiab RNA Isolation yog dab tsi?
Nyob rau hauv DNA cais, ib hom nucleic acid hu ua DNA yog muab rho tawm los ntawm ib tug lom cell, thaum nyob rau hauv RNA cais, ib hom nucleic acid hu ua RNA yog muab rho tawm los ntawm ib tug lom cell. Yog li, qhov no yog qhov sib txawv tseem ceeb ntawm DNA thiab RNA cais. Tsis tas li ntawd, kev cais DNA ib txwm ua nyob ib puag ncig pH 7 txog 8 thaum RNA cais tawm ib txwm ua nyob ib puag ncig pH 4.5.
Cov ntawv qhia hauv qab no nthuav tawm qhov sib txawv ntawm DNA thiab RNA kev cais tawm hauv daim ntawv teev npe rau ib sab ntawm kev sib piv.
Summary – DNA vs RNA Isolation
DNA thiab RNA kev rho tawm yog ob hom kev cais tawm ntawm nucleic acid. DNA cais tawm yog txheej txheem thaum lub sij hawm DNA tau muab rho tawm los ntawm cov kab mob lom. Kev rho tawm RNA yog txheej txheem thaum lub sijhawm RNA raug rho tawm los ntawm lub xov tooj ntawm tes. Yog li, qhov no yog qhov txawv ntawm DNA thiab RNA cais.
