Qhov sib txawv tseem ceeb ntawm genomic DNA thiab plasmid DNA kev rho tawm yog qhov kev rho tawm genomic DNA tsom rau kev rho tawm ntawm genomic DNA thaum lub plasmid DNA cais lub hom phiaj rau kev rho tawm ntawm plasmid DNA ntawm cov kab mob.
Kev cais DNA yog cov txheej txheem tshuaj uas siv los cais DNA ntawm ntau hom lossis los ntawm cov qauv sib txawv. Kev cais DNA yog ib qho tseem ceeb hauv cov txheej txheem hauv qab dej molecular biology, xws li gel electrophoresis, polymerase chain reaction thiab DNA sequencing techniques. Yog li ntawd, kev cais tawm ntawm DNA yog cov txheej txheem tshuaj lom neeg tseem ceeb hauv kev tshawb fawb molecular biological. Raws li lub hom phiaj ntawm kev tshawb fawb, qee zaum nws yog qhov tsim nyog los cais cov genomic DNA. Tsis tas li ntawd, qee qhov kev tshawb fawb tsom rau kev cais plasmid DNA los ntawm cov kab mob. Genomic DNA cais yog cov txheej txheem ntawm kev cais cov genomic DNA los ntawm cov qauv prokaryotic lossis eukaryotic qauv. Cov kauj ruam ntawm kev sib cais txawv raws li hom cell uas DNA cais. Plasmid DNA cais yog cov txheej txheem ntawm kev cais plasmid DNA los ntawm cov kab mob ntawm tes. Nyob rau hauv kev sib piv rau genomic DNA cais, tag nrho cov txheej txheem yog complex nyob rau hauv plasmid DNA cais.
Dab tsi yog Genomic DNA Isolation?
Genomic DNA cais yog cov txheej txheem ntawm kev rho tawm tag nrho genomic DNA ntawm cov kab mob. Cov txheej txheem tshwj xeeb no suav nrog peb qhov xwm txheej tseem ceeb. Lawv yog, cell lysis los yog nuclear lysis, protein degradation los yog proteolysis thiab nag lossis daus ntawm genomic DNA. Cov kauj ruam lysis yuav txawv raws li hom cell. Hauv prokaryotes, vim tias muaj peptidoglycan cell phab ntsa, thawj kauj ruam yuav tsum yog kev tawg ntawm phab ntsa ntawm tes. Ntawm qhov tod tes, hauv eukaryotes, cov kauj ruam lysis cuam tshuam nrog kev tawg ntawm cov plasma membrane thiab cov ntaub so ntswg nuclear tshem tawm DNA mus rau sab nrauv. Nyob rau hauv sib piv, cov kauj ruam tshwj xeeb yog qhov tseem ceeb rau lyse cov nroj tsuag thiab fungal cell phab ntsa.
Daim duab 01: Genomic DNA Isolation
Yog li ntawd, thaum lub sijhawm lysis ua tiav, thaum kawg DNA yuav tuaj rau tus supernatant. Ib txhij, protein degradation kuj tshwm sim nyob rau hauv cov kev daws teeb meem vim qhov sib ntxiv ntawm proteinase K. Cov kauj ruam tom ntej yog cais genomic DNA thiab degraded proteins ntawm ib leeg. Yog li ntawd, cov proteins uas degraded cais tawm los ntawm nag lossis daus uas tso cai rau genomic DNA nyob twj ywm hauv cov supernatant. Tom qab nag lossis daus ntawm cov proteins, genomic DNA tuaj yeem ua rau nag thiab rov muab ncua rau hauv qhov tsis haum kom txog thaum nws xav tau rau qhov kev sim.
Genomic DNA, uas yog linear DNA muaj tag nrho cov ntaub ntawv caj ces ntawm lub cev. Hauv lwm lo lus, genome yog cov khoom siv los ntawm cov kab mob uas muaj sia nyob uas yog lub luag haujlwm rau tag nrho cov txheej txheem thiab kev ua haujlwm ntawm tes. Nws muaj ob qho tib si coding thiab non-coding DNA sequences. Thaum cais genomic DNA, nws suav nrog tag nrho cov genome ntawm cov kab mob.
Plasmid DNA Isolation yog dab tsi?
Plasmid DNA cais tawm yog txheej txheem tshwj xeeb thiab nyuaj ntawm kev cais DNA. Plasmids yog extrachromosomal DNA muaj nyob rau hauv feem ntau cov kab mob hlwb. Lawv yog dormant ncig DNA uas txhawb cov kab mob kom ciaj sia nyob rau hauv ib puag ncig hnyav. Plasmid DNA muaj cov tshuaj tiv thaiv tshwj xeeb uas muab cov txiaj ntsig zoo ntxiv rau cov kab mob xws li tshuaj tua kab mob, tshuaj tiv thaiv kab mob thiab cov tshuaj lom.
Daim duab 02: Plasmid DNA Isolation
Kev cais tawm ntawm Plasmid DNA kuj tseem muaj peb txheej txheem tseem ceeb; cell lysis, proteinolysis thiab DNA nag lossis daus. Txawm hais tias cov txheej txheem biochemical ntawm kev sib cais yog zoo ib yam li cov genomic DNA cais, cov txheej txheem yog ntau dua li nws. Cov txheej txheem cell lysis yog txheej txheem tseem ceeb tshaj plaws hauv cov txheej txheem no. Qhov tseem ceeb tshaj plaws, genomic DNA thiab plasmid DNA yuav tsum tsis txhob sib xyaw ua ke. Yog li ntawd, cov txheej txheem lysis ntau me me sib txuas rau hauv cov txheej txheem plasmid DNA cais tawm. Li no, nyob rau hauv feem ntau plasmid DNA cais cov txheej txheem siv cov tshuaj ntxuav tes; Sodium dodecyl sulfate rau cell lysis.
Dab tsi zoo sib xws ntawm Genomic DNA thiab Plasmid DNA Isolation?
- Genomic DNA thiab Plasmid DNA cais tawm tsom mus rau kev rho tawm DNA ntawm cov kab mob.
- Tsis tas li, ob qho tib si ua raws li cov txheej txheem tag nrho uas suav nrog cell lysis, protein degradation thiab DNA nag lossis daus.
- Tsis tas li ntawd, ua rau DNA ntawm ob txheej txheem tseem ceeb rau cov txheej txheem qis qis.
- Ntxiv mus, ob qho tib si txheej txheem cais suav nrog cov kauj ruam los ntxuav thiab khaws cov DNA raws li cov xwm txheej khaws cia.
- Dhau li ntawm, proteinase K yog siv ob txoj hauv kev los txo cov proteins.
Qhov txawv ntawm Genomic DNA thiab Plasmid DNA Isolation yog dab tsi?
Kev cais DNA Genomic tsom rau kev rho tawm tag nrho cov DNA ntawm cov kab mob hauv lub hom phiaj thaum lub plasmid DNA cais tsom mus rau kev cais cov plasmid DNA los ntawm cov kab mob tshwj xeeb. Yog li, qhov no yog qhov sib txawv tseem ceeb ntawm genomic DNA thiab plasmid DNA cais. Ntxiv mus, lwm qhov sib txawv ntawm genomic DNA thiab plasmid DNA cais yog nyob rau hauv tus txheej txheem. Lub genomic DNA cais yog ib txoj kev tsis tshua muaj kev sib piv rau plasmid DNA cais. Yog li, thaum cais plasmid DNA, nws yuav tsum tau ceev faj kom tsis txhob sib xyaw ntawm genomic thiab plasmid DNA nrog ib leeg.
Cov hauv qab no infograohic muab cov ntsiab lus ntxiv ntawm qhov sib txawv ntawm genomic DNA thiab plasmid DNA cais.
Summary – Genomic DNA vs Plasmid DNA Isolation
kev cais DNA yog txheej txheem tseem ceeb hauv cov txheej txheem biology molecular. Muaj ob hom DNA uas yog genomic DNA thiab plasmid DNA (extra-chromosomal DNA). Raws li qhov yuav tsum tau ua, qee cov txheej txheem tau ua los cais cov genomic DNA thaum qee cov txheej txheem tsom rau kev cais cov plasmid DNA los ntawm cov kab mob. Yog li ntawd, cov kauj ruam koom nrog hauv ob txoj kev sib txawv me ntsis ntawm ib leeg. Txawm li cas los xij, tag nrho cov txheej txheem yog tib yam hauv ob qho tib si cais. Kev rho tawm DNA ntawm ob txheej txheem muaj kev siv loj heev hauv cov txheej txheem qis xws li cloning, gel electrophoresis thiab polymerase chain reactions. Thaum kawg ntawm genomic DNA cais raws tu qauv, tag nrho cov genomic DNA ntawm cov kab mob tuaj yeem raug cais raws li cov khoom kawg thaum kawg ntawm plasmid DNA cais raws tu qauv, plasmid DNA ntawm cov kab mob sib txawv tuaj yeem raug cais raws li cov khoom kawg. Yog li, qhov no yog qhov txawv ntawm genomic DNA thiab plasmid DNA cais.
