He Paepae Hou no ka Hoʻokaʻawale ʻana i ka DNA Wai Kaiapuni — Hoʻolalelale ka Hoʻonohonoho ʻana o Bigefei i ka Noiʻi ʻEpekema

Hoʻokumu ʻia kēia pahu hana ma luna o kahi ʻōnaehana buffer i hoʻomaikaʻi ʻia i hui pū ʻia me nā momi nano magnetic hana kiʻekiʻe. Hoʻopaʻa pono ka DNA genomic i nā hui hana ma ka ʻili o ka momi a hoʻokaʻawale ʻia ma lalo o kahi kahua magnetic waho. Ma hope o nā ʻanuʻu holoi mālie he nui e wehe i nā protein, nā paʻakai, a me nā mea haumia ʻē aʻe, ua hoʻopau hope ʻia ka DNA genomic maʻemaʻe kiʻekiʻe.

Hoʻolālā kūikawā ʻia no nā laʻana wai kaiapuni, unuhi pono ka pahu i ka DNA bacteria i hōʻiliʻili ʻia ma nā membrane kānana, me nā bacteria Gram-negative a me Gram-positive (a hiki i ka 2 × 10⁹ mau cell bacteria no kēlā me kēia membrane kānana hoʻokahi). He kūpono ia me nā ʻōnaehana unuhi waikawa nucleic piha piha no ka hana ʻana i ka throughput kiʻekiʻe. He kūlike ka maikaʻi o ka DNA i unuhi ʻia a hiki ke hoʻohana pololei ʻia no PCR/qPCR, NGS, a me nā noi ʻē aʻe ma hope.

Nā Hiʻohiʻona Huahana

1. Ka hiki ke hoʻokaʻawale i nā bacteria ākea

Hoʻopuka pono i nā bacteria Gram-negative a me Gram-positive mai nā hāpana wai, e uhi ana i nā kaiāulu microbial i loaʻa pinepine i nā wahi wai maʻemaʻe a me ke kai, a hoʻokō i nā koi loiloi like ʻole.

2. Maʻemaʻe kiʻekiʻe a me ka hua kiʻekiʻe

Hāʻawi i ka DNA me ka maʻemaʻe kiʻekiʻe, me ka ʻole o nā mea haumia inhibitory, a me nā hua paʻa e kūpono no nā noi molekala pololei ma hope.

3. Hoʻohālikelike ʻakomi a me ka pono kiʻekiʻe

Hoʻopiha piha ʻia me nā ʻōnaehana hoʻokaʻawale waikawa nucleic automated Bigfish, e kākoʻo ana i ka hana like ʻana o 32 a i ʻole 96 mau laʻana, e hōʻemi nui ana i ka manawa hana a hoʻomaikaʻi i ka pono o ka hale hana.

4. Hana Palekana a maʻalahi hoʻi

ʻAʻohe pono no nā reagents organik ʻawahia e like me ka phenol a i ʻole chloroform, e hōʻemi ana i nā pilikia palekana o ka hale hana. Hoʻopili mua ʻia nā reagents Core i nā pā 96-well, e hōʻemi ana i nā hewa pipetting manual a hoʻomaʻalahi i nā kahe hana.

Nā Mea Hana Kūlike

ʻO Bigfish BFEX-16E

BFEX-32

BFEX-32E

BFEX-96E

Nā Hopena Hoʻokolohua

Ua kānana ʻia kahi hāpana wai muliwai 600 mL ma o kahi membrane, a ua unuhi ʻia ka DNA me ka hoʻohana ʻana i ka Bigfish magnetic bead-based environmental water genomic DNA extraction and purification kit me ka mea hana kūpono. Ma hope mai, ua kālailai ʻia ka DNA i unuhi ʻia e ka agarose gel electrophoresis.