Tekanyetso e Ncha ea ho Ntša DNA ea Metsi Tikolohong — Bigefei Sequencing e Potlakisa Lipatlisiso tsa Saense

Sete ena e thehiloe tsamaisong e ntlafalitsoeng ea buffer e kopantsoeng le li-nano magnetic beads tse sebetsang hantle. DNA ea genomic e tlamahane ka ho khetheha le lihlopha tse sebetsang holim'a li-beads 'me e aroloa tlas'a matla a makenete a kantle. Kamora mehato e mengata ea ho hlatsoa ka bonolo ho tlosa liprotheine, letsoai le litšila tse ling, DNA ea genomic e hloekileng haholo e qetella e le sieo.

E etselitsoe ka ho khetheha disampole tsa metsi a tikoloho, kit ena e ntša DNA ea baktheria e bokelletsoeng ka katleho lera la sefe, ho kenyeletsoa le libaktheria tsa Gram-negative le Gram-positive (ho fihlela liseleng tsa baktheria tse 2 × 10⁹ ka lera le le leng la sefe). E lumellana le litsamaiso tsa ho ntša nucleic acid tse iketsang ka botlalo bakeng sa ts'ebetso e phahameng ea tlhahiso. DNA e ntšitsoeng e na le boleng bo tsitsitseng 'me e ka sebelisoa ka kotloloho bakeng sa PCR/qPCR, NGS, le lits'ebetso tse ling tse latelang.

Likarolo tsa Sehlahisoa

1. Bokhoni ba ho Ntša Baktheria ea Broad-Spectrum

E ntša libaktheria tse nang le Gram-negative le Gram-positive ka katleho lisampoleng tsa metsi, e akaretsa metse ea likokoana-hloko tse fumanehang hangata metsing a hloekileng le a leoatle, 'me e fihlela litlhoko tse fapaneng tsa tlhahlobo.

2. Bohloeki bo Phahameng le Tlhahiso e Phahameng

E fana ka DNA ka bohloeki bo phahameng, e se nang litšila tse thibelang, le chai e tsitsitseng e loketseng ts'ebeliso e tobileng ea limolek'hule.

3. Ho lumellana ka boiketsetso le ka bokgoni bo phahameng

E lumellana ka botlalo le litsamaiso tsa ho ntša nucleic acid tse iketsahallang tsa Bigfish, e tšehetsang ts'ebetso e ts'oanang ea lisampole tse 32 kapa tse 96, e fokotsang nako ea ts'ebetso haholo le ho ntlafatsa katleho ea laboratori.

4. Ts'ebetso e Sireletsehileng le e Bonolo ho Basebelisi

Ha ho hlokahale di-reagent tse chefo tsa tlhaho tse kang phenol kapa chloroform, e leng se fokotsang dikotsi tsa polokeho ya laboratori. Di-reagent tsa mantlha di kentswe ka hara di-plate tsa di-well tse 96, ho fokotsa diphoso tsa ho kenya diphaephe ka letsoho le ho nolofatsa mesebetsi.

Lisebelisoa tse Lumellanang

Tlhapi e Kholo BFEX-16E

BFEX-32

BFEX-32E

BFEX-96E

Liphetho tsa Liteko

Sampole ea metsi a noka ea 600 mL e ile ea sefshoa ka lera, 'me DNA ea ntšoa ho sebelisoa sesebelisoa sa Bigfish magnetic bead sa ho ntša le ho hloekisa genomic DNA hammoho le sesebelisoa se lumellanang. DNA e ntšitsoeng e ile ea hlahlojoa hamorao ke agarose gel electrophoresis.