ʻO TIANamp Virus RNA Kit

A-1 Hōʻemi haʻahaʻa o nā hunaola a i ʻole virus i ka hāpana hoʻomaka
—E hoʻonui i ka nui o nā hunaola a i ʻole nā ​​virus.

A-2 Lysis lawa ʻole o nā laʻana
-ʻAʻole i hui maikaʻi ʻia nā laʻana me ka buffer lysis. Manaʻo ʻia e kāwili pono ʻia e ka pulse-vortexing no 1-2 mau manawa.
—Ua lawa ʻole ka lysis pūnaewele e hoʻoliʻiliʻi ʻia e ka hana o proteinase K.
—No ka lawa ʻole o ka lysis pūnaewele a i ʻole ka hōʻino protein ʻana ma muli o ka lawa ʻole o ka manawa ʻauʻau mehana. 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.

Ua lawe ʻia ka genomic DNA mai nā puke like ʻole o nā laʻana koko kanaka āpau e TIANamp Blood DNA Kit. Penei nā hopena. Ua helu ʻia nā hopena ma ke ʻano he kūmole wale nō, nā hopena o ka unuhi maoli ʻana ma muli o nā kūlana o nā laʻana.

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).