puas yog lub centrifuge cais cov DNA ntawm cov protein
Puas yog lub Centrifuge cais DNA ntawm cov Proteins?
Kev Taw Qhia
Kev siv centrifugation yog ib txoj kev tseem ceeb uas siv dav hauv kev tshawb fawb thiab cov chaw kuaj mob rau kev sib cais ntau yam khoom ntawm cov qauv raws li lawv qhov ceev. Ib qho kev siv ntau ntawm cov txheej txheem no yog kev cais DNA thiab cov protein ntawm cov qauv kab mob. Hauv tsab xov xwm no, peb yuav tshawb nrhiav seb lub centrifuge puas muaj peev xwm cais DNA ntawm cov protein thiab nkag mus rau hauv ntau txoj kev sib txawv uas siv los ua kom tiav kev sib cais zoo.
Nkag Siab Txog Kev Siv Centrifugation
Ua ntej yuav tham txog cov ntsiab lus tshwj xeeb ntawm kev cais DNA ntawm cov protein siv lub centrifuge, nws yog ib qho tseem ceeb kom nkag siab txog cov ntsiab cai tseem ceeb ntawm centrifugation. Lub centrifuge ua haujlwm raws li lub hauv paus ntsiab lus ntawm kev tso dej thiab siv kev sib hloov ceev ceev los tsim lub zog centrifugal muaj zog. Lub zog no ua rau cov khoom me me hauv cov qauv sib cais raws li lawv qhov ceev, nrog rau cov khoom me me dua txav mus rau hauv qab ntawm lub raj qauv. Yog li ntawd, cov khoom me me dua feem ntau nyob rau sab saum toj.
Qhov Kev Sib Tw ntawm Kev Sib Cais DNA thiab Cov Protein
Thaum nws los txog rau kev cais DNA thiab cov protein, nws yog ib qho kev cov nyom vim tias ob qho tib si molecule muaj qhov hnyav molecular zoo sib xws. Yog tsis muaj txoj kev sib cais zoo, nws nyuaj rau kom tau txais cov qauv DNA lossis protein ntshiab. Txawm li cas los xij, centrifugation tuaj yeem pab hauv cov txheej txheem sib cais, tab sis nws yuav tsis txaus ntawm nws tus kheej.
1. Kev Sib Tw Centrifugation: Kev Sib Cais Pib
2. Ultracentrifugation: Kev Sib Cais Zoo Dua
3. Kev Siv Centrifugation Gradient Density: Txhim Kho Cov Txheej Txheem
4. Qhov Tseem Ceeb ntawm Kev Xaiv Buffer
5. Kev Centrifugation piv rau Lwm Cov Txheej Txheem Sib Cais
Kev Sib Tw Centrifugation Sib Txawv: Kev Sib Cais Thaum Pib
Kev sib txawv centrifugation yog thawj kauj ruam hauv kev cais DNA ntawm cov protein. Txoj kev no siv cov centrifugation ceev sib txawv kom ua tiav kev sib cais dav dav ntawm cov khoom cellular. Thaum lub sijhawm ua cov txheej txheem no, cov cell homogenized lossis cov qauv ntaub so ntswg raug rau centrifugation qeeb qeeb, feem ntau nyob ib puag ncig 1000-5000 revolutions ib feeb (rpm). Qhov kev sib hloov qeeb qeeb no ua rau kev sib cais ntawm cov nuclei thiab cov cell intact, ua rau muaj kev faib ua feem tsis zoo. Txawm li cas los xij, nws tsis muab kev sib cais tag nrho ntawm DNA thiab cov protein.
Ultracentrifugation: Kev Sib Cais Zoo Dua
Yuav kom kov yeej qhov kev txwv ntawm kev sib txawv centrifugation, ultracentrifugation nkag mus rau hauv qhov chaw. Ultracentrifuges yog cov centrifuges ceev ceev uas muaj peev xwm ncav cuag qhov ceev txog li 100,000 rpm. Los ntawm kev siv cov feem pua ntawm cov khoom siv raw los ntawm kev pib qis qis centrifugation mus rau ultracentrifugation, qib siab dua ntawm kev sib cais ntawm DNA thiab cov protein tuaj yeem ua tiav. Cov DNA molecules hnyav sediment mus rau hauv qab ntawm lub raj, tsim cov pellet pom, thaum cov protein tseem nyob hauv supernatant.
Kev Siv Centrifugation Gradient Density: Txhim Kho Cov Txheej Txheem
Yuav kom txhim kho qhov kev sib cais ntawm DNA thiab cov protein, kev siv density gradient centrifugation. Hauv cov txheej txheem no, ib qho density gradient medium, xws li sucrose lossis cesium chloride, tau npaj. Cov khoom no muaj ntau yam densities, nce ntawm sab saum toj mus rau hauv qab ntawm lub raj centrifuge. Cov fraction crude tau los ntawm cov kauj ruam dhau los yog ua tib zoo layered rau saum toj ntawm qhov density gradient thiab raug rau ultracentrifugation dua ib zaug. Cov DNA thiab cov protein sib cais raws li lawv cov densities thiab tsim cov bands sib txawv hauv qhov gradient. Fractionation ntawm qhov gradient tso cai rau kev rov qab tau cov DNA thiab cov protein qauv ntshiab.
Qhov Tseem Ceeb ntawm Kev Xaiv Buffer
Cov buffer siv thaum lub sijhawm centrifugation ua lub luag haujlwm tseem ceeb hauv kev ua tiav kev sib cais zoo. Nws cuam tshuam rau kev ruaj khov thiab kev ncaj ncees ntawm cov molecules raug sib cais. Rau DNA, Tris-EDTA buffer nrog pH ntawm 8.0 feem ntau yog siv vim nws muaj peev xwm tswj hwm DNA ruaj khov. Ntawm qhov tod tes, cov protein xav tau cov buffers nrog cov pH tshwj xeeb, feem ntau nyob ib puag ncig 7.0, los tiv thaiv denaturation thiab tswj hwm lawv cov qauv ib txwm muaj. Kev xaiv cov buffer tsim nyog ua kom ntseeg tau tias kev sib cais zoo ntawm DNA thiab cov protein yam tsis muaj kev cuam tshuam rau lawv qhov zoo.
Centrifugation piv rau lwm cov txheej txheem sib cais
Txawm hais tias centrifugation yog ib txoj kev siv dav hauv kev sib cais DNA ntawm cov protein, nws yog ib qho tseem ceeb kom lees paub tias nws yuav tsis yog tib txoj kev siv. Lwm cov txheej txheem, xws li chromatography, electrophoresis, thiab precipitation, tuaj yeem ua tiav centrifugation kom ua tiav kev sib cais zoo dua. Txhua txoj kev muaj nws tus kheej qhov zoo thiab kev txwv, thiab cov kws tshawb fawb feem ntau ua ke ntau txoj hauv kev kom tau txais qhov huv si thiab rov qab tau ntawm DNA thiab cov protein.
Xaus lus
Xaus lus, thaum lub centrifuge yeej muaj txiaj ntsig zoo rau kev cais DNA ntawm cov protein, nws ua haujlwm zoo tshaj plaws ua ke nrog lwm cov txheej txheem. Kev sib txawv centrifugation, ultracentrifugation, thiab density gradient centrifugation tsim cov kauj ruam sib cais uas pab tau kom tau txais cov qauv huv dua. Tsis tas li ntawd, kev xaiv cov buffers tsim nyog thiab kev xav txog lwm cov txheej txheem sib cais ntxiv txhim kho qhov ua tau zoo ntawm kev sib cais DNA-protein.
.