He aha ke ʻano hana no ka hoʻokaʻawale ʻana a me ka hoʻomaʻemaʻe ʻana i nā waikawa nucleic?

Nā waikawa nukleika, me ka DNA a me ka RNA, he mau biomolecules koʻikoʻi e pāʻani ana i kahi kuleana koʻikoʻi i ka genetics, molecular biology, a me ka biotechnology. ʻO ka hiki ke hoʻokaʻawale a hoʻomaʻemaʻe i kēia mau waikawa nucleic he mea nui ia no nā ʻano noi like ʻole, me ka cloning, sequencing, a me ka nānā ʻana i ka gene expression. Hoʻopuni nā ʻōnaehana hoʻomaʻemaʻe waikawa nucleic i nā ʻano hana i hoʻolālā ʻia e unuhi a hoʻomaʻemaʻe i nā waikawa nucleic mai nā laʻana olaola. Ke ʻimi nei kēia ʻatikala i nā ʻano hana no ka hoʻokaʻawale ʻana a me ka hoʻomaʻemaʻe ʻana i nā waikawa nucleic a hōʻike i ke koʻikoʻi o kēia mau ʻōnaehana i ka noiʻi ʻepekema hou.

Ke hoʻomaopopo nei i ka hoʻomaʻemaʻe ʻana o ka waikawa nucleic

ʻO ka hoʻomaʻemaʻe ʻana o ka waikawa nucleic e pili ana i ka unuhi ʻana o ka DNA a i ʻole RNA mai nā cell a i ʻole nā ​​​​ʻiʻo, a ukali ʻia e ka wehe ʻana i nā mea haumia e like me nā protein, lipids, a me nā ʻōpala cellular ʻē aʻe. He mea koʻikoʻi ka maʻemaʻe a me ka pono o nā waikawa nucleic i hoʻokaʻawale ʻia no nā noi ma hope, no ka mea hiki i nā mea haumia ke kāohi i nā hopena enzymatic a hoʻopilikia i ka pololei o nā hopena hoʻokolohua.

Nā ʻano maʻamau no ka hoʻokaʻawale ʻana a me ka hoʻomaʻemaʻe ʻana i ka waikawa nucleic

Ka unuhi ʻana o ka Phenol-chloroform:Hoʻohana kēia ʻano hana kuʻuna i nā mea hoʻoheheʻe organik e hoʻokaʻawale i nā waikawa nucleic mai nā protein a me nā ʻāpana cellular ʻē aʻe. Hoʻohui ʻia ka laʻana me ka phenol a me ka chloroform, e hoʻokaʻawale ana i nā waikawa nucleic i loko o ka pae wai, ʻoiai e noho ana nā protein i ka pae organik. Ma hope o ka centrifugation, hōʻiliʻili ʻia ka pae wai e loaʻa ana nā waikawa nucleic a hoʻoheheʻe ʻia me ka ethanol.

Nā ʻano hana i hoʻokumu ʻia i ka gel silica:Hoʻohana nui ʻia nā membrane silica gel i nā pahu hoʻomaʻemaʻe waikawa nucleic kalepa. ʻO ke kumumanaʻo o kēia ʻano hana, ʻo ia ka hoʻopaʻa ʻana o nā waikawa nucleic i ka silica gel ma nā paʻakai kiʻekiʻe. Ma hope o ka hoʻopaʻa ʻana, holoi ʻia nā mea haumia, a laila hoʻokahe ʻia nā waikawa nucleic me kahi buffer paʻakai haʻahaʻa a i ʻole ka wai. Makemake ʻia kēia ʻano hana no ka mea he wikiwiki, kūpono, a loaʻa nā waikawa nucleic maʻemaʻe kiʻekiʻe.

Hoʻomaʻemaʻe ʻana i nā momi magnetic:Hoʻohana kēia ʻenehana i nā momi magnetic i uhi ʻia me nā mea hoʻopaʻa waikawa nucleic. Ke hui ʻia kahi laʻana me nā momi magnetic, hoʻopili nā waikawa nucleic ma luna o ka ʻili o nā momi. A laila hoʻokaʻawale ʻia nā momi mai ka hopena me ka hoʻohana ʻana i kahi magnet, a laila e wehe ana i nā mea haumia. He ʻano hana like ʻole kēia a hiki ke hana ʻakomi ʻia, e kūpono ai no nā noi throughput kiʻekiʻe.

Kromatografi kolamu:ʻO kēia ʻano hana e pili ana i ka hoʻoili ʻana i kahi hāpana ma o kahi kolamu chromatographic i hoʻopiha ʻia me kahi pae paʻa e mālama koho ana i nā waikawa nucleic. Hiki ke hoʻohana ʻia nā ʻano kolamu like ʻole, me nā mea i hoʻokumu ʻia ma ka hoʻokaʻawale ʻana i ka nui a i ʻole nā ​​​​kumumanaʻo hoʻololi ion. Hoʻokuʻu ʻia nā waikawa nucleic mai ke kolamu, e hopena ana i kahi hāpana i hoʻomaʻemaʻe ʻia.

Nā ʻano hana enzymatic:Hiki ke hoʻohana ʻia nā ʻano hana enzymatic, e like me nā mea e hoʻohana ana iā DNase a i ʻole RNase, e hoʻohaʻahaʻa koho i nā waikawa nucleic a i ʻole nā ​​​​mea haumia i makemake ʻole ʻia. He kūpono loa kēia ʻano hana i ka wā e hana ai i nā laʻana paʻakikī, e like me nā mea i loaʻa ka DNA a me ka RNA.

I ka hopena

ʻO ka hoʻokaʻawale ʻana a me ka hoʻomaʻemaʻe ʻana i ka waikawa nucleic he mau ʻanuʻu koʻikoʻi i ka noiʻi a me nā noi o ka biology molekala.Nā ʻōnaehana hoʻomaʻemaʻe waikawa nukleikahāʻawi i nā mea noiʻi i nā ʻano hana like ʻole e loaʻa ai nā waikawa nucleic kiʻekiʻe e kūpono no nā noi ma hope. Inā paha e hoʻohana ana i ka unuhi phenol-chloroform kuʻuna a i ʻole nā ​​​​ʻano hana hou e like me ka silica gel a i ʻole ka hoʻomaʻemaʻe magnetic bead-based, ʻo ke koho ʻana o ke ʻano hana e pili ana i nā koi kikoʻī o ka hoʻokolohua a me ke ʻano o ka laʻana. Me nā holomua ʻenehana, ua ulu mau kēia mau ʻōnaehana hoʻomaʻemaʻe, e hoʻomaikaʻi ana i ka pono, ka wikiwiki, a me ka hilinaʻi, a hoʻomaikaʻi hope loa i nā hiki o nā mea noiʻi ma ke kahua o ka biology molecular.