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Indlela yeMagnetic Bead isombulula ngempumelelo imingeni ekukhutshweni kweDNA yamanzi kwiNdalo
Kwimimandla efana nophando lwe-microbiology yendalo kunye nokubeka esweni ungcoliseko lwamanzi, ukukhutshwa kwe-DNA ye-genomic esemgangathweni ophezulu kuyimfuneko ebalulekileyo kwizicelo ezisezantsi kuquka i-PCR/qPCR kunye ne-next-generation sequencing (NGS). Nangona kunjalo, iisampulu zamanzi endawo ziyinkimbinkimbi kakhulu, ziqulathe amaqela ahlukeneyo eentsholongwane, iintlobo ezinzima uku-lyse ezifana ne-Gram-positive bacteria, kunye nemingeni yexesha elide enxulumene neendlela zemveli zokukhupha-ezifana nokusetyenziswa kwee-reagents ezinobuthi kunye neenkqubo ezinzima-eziye zaphazamisa abaphandi rhoqo.
Ngoku, iBigfish Sequencing izisa iBFMP24R Magnetic Bead-Based Environmental Water Genomic DNA Extraction and Purification Kit, ebonelela ngesisombululo esipheleleyo kwezi ngxaki ngokusebenzisa ubuchwepheshe obutsha kunye noyilo olusebenziseka lula.
Isishwankathelo seMveliso
Le khithi isekelwe kwinkqubo ye-buffer elungiselelwe kakuhle edityaniswe nee-nano magnetic beads ezisebenza kakuhle. I-Genomic DNA ibopha ngokukodwa kumaqela asebenzayo kumphezulu wee-beads kwaye yahlulwe phantsi kwentsimi yemagnethi yangaphandle. Emva kwamanyathelo amaninzi okuhlamba ngobunono ukususa iiproteni, iityuwa, kunye nezinye izinto ezingcolileyo, i-genomic DNA ecocekileyo kakhulu ekugqibeleni ayifumaneki.
Yenzelwe ngokukodwa iisampulu zamanzi okusingqongileyo, le khithi ikhupha ngempumelelo i-DNA yebhaktiriya eqokelelwe kwiimembrane zesihluzo, kuquka iibhaktiriya zeGram-negative kunye neGram-positive (ukuya kuthi ga kwiiseli zebhaktiriya ezi-2 × 10⁹ kwimembrane yesihluzo esinye). Ihambelana neenkqubo zokukhupha i-nucleic acid ezizenzekelayo ngokupheleleyo ukuze kusetyenzwe nge-high-throughput. I-DNA ekhutshiweyo inomgangatho ofanayo kwaye ingasetyenziselwa ngokuthe ngqo kwi-PCR/qPCR, NGS, nakwezinye iinkqubo ezisezantsi.
Iimpawu zeMveliso
1. Amandla okuKhupha iiBhaktheriya eziBanzi
Ikhupha iintsholongwane zeGram-negative kunye neGram-positive ngokufanelekileyo kwiisampuli zamanzi, igubungela uluntu lweentsholongwane olufumaneka rhoqo kwiindawo zamanzi acocekileyo naselwandle, kwaye ihlangabezana neemfuno ezahlukeneyo zohlalutyo.
2. Ubumsulwa obuphezulu kunye nemveliso ephezulu
Inika i-DNA ngobunyulu obuphezulu, ayinazo izinto ezithintela ukungcola, kwaye ivelisa imveliso ezinzileyo efanelekileyo kwiindlela ezisetyenziswa ngqo kwiimolekyuli.
3. Ukuhambelana okuzenzakalelayoyo nokusebenza kakuhle
Ihambelana ngokupheleleyo neenkqubo zokukhupha i-nucleic acid ezizenzekelayo zeBigfish, ezixhasa ukucutshungulwa ngaxeshanye kweesampulu ezingama-32 okanye ezingama-96, zinciphisa kakhulu ixesha lokucubungula kwaye ziphucula ukusebenza kakuhle kwelebhu.
4. Ukusebenza okuKhuselekileyo nokuLungelelanisayo
Akukho mfuneko yee-reagents ezinobungozi eziphilayo ezifana ne-phenol okanye i-chloroform, nto leyo enciphisa umngcipheko wokhuseleko lwelabhoratri. Ii-reagents eziphambili zipakishwa kwangaphambili kwiipleyiti ezingama-96-well, nto leyo enciphisa iimpazamo zokufaka imibhobho ngesandla kwaye yenza kube lula ukuhamba komsebenzi.
Izixhobo Ezihambelanayo
Intlanzi enkulu iBFEX-16E
I-BFEX-32
I-BFEX-32E
I-BFEX-96E
Iziphumo zovavanyo
Isampulu yamanzi omlambo eyi-600 mL yahluzwa nge-membrane, kwaye i-DNA yakhutshwa kusetyenziswa i-Bigfish magnetic bead-based environmental water genomic DNA extraction and purification kit kunye nesixhobo esihambelanayo. I-DNA ekhutshiweyo yahlaziywa kamva yi-agarose gel electrophoresis.
M: Uphawu1, 2: Iisampulu zamanzi omlambo
Iinkcukacha zeMveliso
Ixesha leposi: Disemba-18-2025
Izicwangciso zabucala