Isilinganiso Esisha Sokukhishwa Kwe-DNA Yamanzi Emvelo — Ukulandelelana Kwe-Bigefei Kusheshisa Ucwaningo Lwesayensi

Le khithi isekelwe ohlelweni lwe-buffer oluthuthukisiwe oluhlanganiswe nama-nano magnetic beads asebenza kahle kakhulu. I-Genomic DNA ibopha ngqo amaqembu asebenzayo ebusweni be-beads futhi ihlukaniswa ngaphansi kwensimu yamagnetic yangaphandle. Ngemva kwezinyathelo eziningi zokuhlanza ngobumnene ukususa amaprotheni, usawoti, kanye nokunye ukungcola, i-genomic DNA emsulwa kakhulu ekugcineni ayitholakali.

Iklanyelwe ngqo amasampula amanzi emvelo, le khithi ikhipha ngempumelelo i-DNA yamagciwane eqoqwe kuma-membrane esihlungi, kufaka phakathi ama-bacteria angenayo i-Gram kanye ne-Gram-positive (afinyelela kumaseli amabhaktheriya angu-2 × 10⁹ nge-membrane yesihlungi esisodwa). Ihambisana nezinhlelo zokukhipha i-nucleic acid ezizenzakalelayo ngokuphelele zokucubungula okuphezulu. I-DNA ekhishwe inekhwalithi efanayo futhi ingasetshenziswa ngqo kwi-PCR/qPCR, NGS, nakwezinye izinhlelo zokusebenza ezingezansi.

Izici Zomkhiqizo

1. Amandla Okukhipha Amagciwane Ane-Broad-Spectrum

Ikhipha ngempumelelo amabhaktheriya e-Gram-negative kanye ne-Gram-positive kumasampula amanzi, imboze imiphakathi yamagciwane avame ukutholakala ezindaweni zamanzi ahlanzekile kanye nezasolwandle, futhi ihlangabezana nezidingo ezahlukahlukene zokuhlaziya.

2. Ukuhlanzeka Okuphezulu kanye Nesivuno Esiphezulu

Iletha i-DNA ngobumsulwa obuphezulu, ayinazo izinto ezivimbela ukungcola, kanye nemikhiqizo eqinile efanele ukusetshenziswa ngqo kwama-molecule.

3. Ukuhambisana okuzenzakalelayo nokusebenza kahle kakhulu

Ihambisana ngokugcwele nezinhlelo zokukhipha i-nucleic acid ezenzakalelayo ze-Bigfish, isekela ukucutshungulwa ngasikhathi sinye kwamasampula angu-32 noma angu-96, okunciphisa kakhulu isikhathi sokucubungula nokuthuthukisa ukusebenza kahle kwelabhorethri.

4. Ukusebenza Okuphephile Nokulula Ukukusebenzisa

Asikho isidingo sama-reagent anobuthi e-organic njenge-phenol noma i-chloroform, okunciphisa izingozi zokuphepha kwelabhorethri. Ama-reagent ayinhloko afakwa kusengaphambili emapuletini angu-96-well, okunciphisa amaphutha okufaka amapayipi ngesandla futhi kwenza kube lula ukuhamba komsebenzi.

Izinsimbi Ezihambisanayo

Inhlanzi Enkulu i-BFEX-16E

I-BFEX-32

I-BFEX-32E

I-BFEX-96E

Imiphumela Yokuhlola

Isampula yamanzi omfula engama-600 mL yahlungwa nge-membrane, kwathi i-DNA yakhishwa kusetshenziswa i-Bigfish magnetic bead-based environmental water genomic DNA extraction and purification kit kanye nethuluzi elihambisanayo. I-DNA ekhishwe yahlaziywa kamuva yi-agarose gel electrophoresis.