Hiki ke hoʻohana pololei ʻia ʻo DNA i hoʻomaʻemaʻe ʻia i nā ʻano hana like ʻole, e like me ka digestive enzymatic, PCR, kūkulu hale waihona puke, South blot a me nā hoʻokolohua ʻē aʻe.
■ Ka hoʻohaʻahaʻa ʻana i ka mīkini: hiki ke hoʻopau pono ʻia nā hana deparaffinization a me ka ʻoki ʻana me ka waiho wale ʻana i nā ʻāpana paraffin i hana ʻole ʻia i loko o ka pahu pahu reagent a laila hoʻohui i ka Sula Oil a me Proteinase K.
■ Nā hopena kūpono: Loaʻa ka genome DNA mai ka RNA a me ka protein protein, a hiki ke hoʻohana pono ʻia no PCR a i ʻole fluorescence PCR nui.
■ Palekana a palekana ʻole: ʻAʻole pono nā mea hoʻoheheʻe i nā mea kino i ke kino o ke kanaka e like me phenol chloroform.
Hiki ke hoʻonohonoho pono ʻia nā huahana āpau no ODM / OEM. No nā kikoʻī,e kaomi i ka lawelawe i hoʻopilikino ʻia (ODM / OEM)
Ua hōʻike ʻia nā hopena no nā laʻana o ka nui o ka nui, ua hoʻohālikelike ʻia nā hua i kāʻili ʻia o nā ʻano ʻelua; no nā laʻana me ka leo waena a me ka liʻiliʻi, ʻo ka hua o ka hana deparaffinization ʻanuʻu hoʻokahi (hana me TGuide) ʻoi aku ka nui ma mua o ka hana deparaffinization maʻamau. No laila, ʻo ka hana deparaffinization ʻanuʻu wale nō ʻaʻole maʻalahi i ka hana, hoʻemi i ka makaʻu, akā hoʻomaikaʻi pū kekahi i ka pono o ka unuhi ʻana. Kaha: ʻO kahi hāpana nui: aia ka ʻāpana o ka ʻāpana ma waena o 2.5-4 cm2 a aia ka mānoanoa ma waena o 5-20 μm. ʻO kahi laʻana o ka leo waena: ʻo ka ʻāpana o ka mahele ma waena o 1.5-2.5 cm2 a aia ka mānoanoa ma waena o 5-20 μm. ʻO kahi laʻana o ka leo liʻiliʻi: aia ka ʻāpana mahele ma waena o 0.2-1.5 cm2 a aia ka mānoanoa ma waena o 5-20 μm. |
A-1 Haʻahaʻaha haʻahaʻa o nā hunaola a i ʻole virus i ka hōʻailona hoʻomaka - E hoʻonui i ka hoʻonui ʻia o nā hunaola a i ʻole nā virus.
A-2 lawa ʻole ka lisis o nā laʻana - ʻAʻole i huikau hui ʻia nā laʻana me ka buffer lysis. Manaʻo ʻia e kāwili pono ʻia e ka pulse-vortexing no 1-2 mau manawa. —No ka lawa ʻole o ka lysis pūnaewele i hoʻoliʻiliʻi ʻia e ka hana i hoʻoliʻiliʻi o proteinase K. ʻLelo ʻia e ʻokiʻoki i ke kiko i nā ʻāpana liʻiliʻi a hoʻolōʻihi i ka manawa ʻauʻau e lawe i nā koena āpau i ka lysate.
A-3 lawa ʻole ka adsorption DNA. —No ka ethanol a i ʻole ka pākēneka haʻahaʻa ma kahi o 100% ethanol i hoʻohui ʻia ma mua o ka hoʻoili ʻia ʻana o ka lysate i ke kolamu wili.
A-4 ʻOi aku ka waiwai o ka pH o ka buffer elution. — Hoʻoponopono i ka pH ma waena o 8.0-8.3.
Ethanol koena i ka eluent.
— Aia ke koena holoi pā pale PW ma nā eluent. Hiki ke hemo ʻia ka ethanol e ka centrifuging ʻana i ke kolamu wili no 3-5 min, a laila kau i ke ana wela a i ʻole 50 ℃ incubator no 1-2 min.
A-1 ʻAʻole hou ka laʻana. —E unuhi i kahi hāpana DNA kūpono e like me ke kaohi e hoʻoholo inā ua hoʻohaʻahaʻa ʻia ka DNA.
A-2 lapaʻau kūpono ʻole. —Noho ʻia e ka wili nui ʻana o ka nitrogen wai, hoʻāʻo hou, a i ʻole ka nui o nā hāpana.
Pono e hoʻololi i nā pretreatment no nā laʻana like ʻole. No nā mea kanu mea kanu, e wili pono i ka nitrogen wai. No nā laʻana holoholona, homogenate a wili maikaʻi paha i ka nitrogen wai. No nā laʻana me nā paia paʻa e paʻakikī e haki, e like me G + bacteria a me ka hū, ʻōlelo ʻia e hoʻohana i ka lysozyme, lyticase a i ʻole nā ʻano hana mechanical e haki i nā paia o ka pūnaewele.
4992201/4992202 Plant Genomic DNA Kit apono i ke kolamu hana-e koi ana i ka chloroform no ka unuhi. Kūpono ia no nā mea kanu mea kanu like ʻole, a me ka pauka maloʻo maloʻo. ʻO ka Hi-DNAsecure Plant Kit kahi kolamu hoʻi, akā ʻaʻohe pono o ka phenol / chloroform extraction, e palekana ana a ʻaʻohe mea ʻawahia. Kūpono ia no nā mea kanu me nā polysaccharides kiʻekiʻe a me nā ʻike polyphenol. 4992709/4992710 DNAquick Plant System apono i kahi hana wai-pili. ʻAʻole pono ʻo Phenol / chloroform extraction pū kekahi. He maʻalahi a wikiwiki hoʻi ke kaʻina hana hoʻomaʻemaʻe me ka palena ʻole no nā laʻana o ka laʻana, no laila hiki i nā mea hoʻohana ke hoʻomaʻalahi i ka nui e like me nā koi hoʻokolohua. Hiki ke kiʻi ʻia ka nui o nā ʻāpana gDNA me nā hua kiʻekiʻe.
Hiki ke hoʻokō ʻia ke kāʻili ʻana o ka DNA kūpē ʻana i ke koko me ka hoʻohana ʻana i nā reagents i hāʻawi ʻia i loko o kēia mau pahu ʻelua ma o ka hoʻololi ʻana i ka protocol i ke kuhikuhi kikoʻī no ka wehe ʻana o ke koko koko. Hiki ke hoʻopuka i ke kope palupalu o ke kōkō DNA kāʻili i ke kaʻina koi ma ke noi ʻana.
Wehe i ka hāpana hou me 1 ml PBS, saline maʻamau a i ʻole buffer TE. Hoʻopiha piha i ka hāpana e kahi homogenizer a hōʻiliʻili i ka piʻi i lalo o kahi o ka pahu e centrifuging. E hoʻolei i ka mea kupanaha, a resuspend i ka piʻi ʻana me 200 μl buffer GA. Hiki ke hana i ka hoʻomaʻemaʻe DNA aʻe e like me ke aʻo.
No ka hoʻomaʻemaʻe o gDNA i ka plasma, serum a me nā laʻana wai o ke kino, paipai ʻia ʻo TIANamp Micro DNA Kit. No ka hoʻomaʻemaʻe ʻana i ka maʻi gDNA mai nā mea serum / plasma, makemake ʻia ʻo TIANamp Virus DNA / RNA Kit. No ka hoʻomaʻemaʻe ʻana i ka gDNA bacteria mai nā serum a me nā hāpana plasma, paipai ʻia ʻo TIANamp Bacteria DNA Kit (pono e hoʻokomo ʻia ka lysozyme no ka maʻi maikaʻi). No nā laʻana kuha, paipai ʻia ʻo Hi-Swab DNA Kit a me TIANamp Bacteria DNA Kit.
Kākoʻo ʻia ʻo DNAsecure Plant Kit a i ʻole DNAquick Plant System no ka lawe ʻana i ka genome fungal. No ka hemo genome hū, koi ʻia ʻo TIANamp Yeast DNA Kit (e hoʻomākaukau pono ʻia ka lyticase).
Mai kona hoʻokumu ʻia ʻana, ke hoʻomohala nei kā mākou hale hana i nā huahana papa honua mua me ka hoʻopaʻa ʻana i ke ʻano
o ka maikaʻi ma mua. Ua loaʻa i kā mākou huahana ka maikaʻi maikaʻi loa i ka ʻoihana a me ka waiwaiabletrusty i waena o nā mea kūʻai aku a kahiko.