Loaʻa iā mākou i kēia manawa kekahi o nā mīkini hana kiʻekiʻe loa, nā ʻenehana loea a me nā limahana, manaʻo ʻia nā ʻōnaehana hoʻokele maikaʻi a me kahi pūʻulu kūʻai akamai aloha mua / ma hope o ke kūʻai ʻana i ke kumu kūʻai no ka Rapid and High Yeild Virus Total Rna Isolation Extraction Purification Kit Nucleic Acid Detection Kit, "Making the Products of Large Quality" ʻo ia ke kumu mau loa o kā mākou ʻoihana.Hana mākou i ka hoʻoikaika mau ʻana e ʻike i ka pahuhopu o "We Will Always Hold in Pace along with the Time".
Loaʻa iā mākou i kēia manawa kekahi o nā mīkini hana kiʻekiʻe loa, nā ʻenehana loea a me nā limahana, i manaʻo ʻia nā ʻōnaehana hoʻokele maikaʻi a me kahi hui kūʻai akamai aloha ma mua / ma hope o ke kūʻai aku i kākoʻo noChina Universal Rna DNA Reagents a me Automatic Nucleic Acid Purification, I mea e hoʻokō ai i kā mākou pahuhopu o "ka mea kūʻai mua a me ka pōmaikaʻi like" i ka hui pū ʻana, hoʻokumu mākou i kahi hui ʻenekinia ʻoihana a me kahi hui kūʻai aku e hoʻolako i ka lawelawe maikaʻi loa e hoʻokō i nā koi o kā mākou mea kūʻai.Welina mai iā ʻoe e hui pū me mākou a hui pū me mākou.ʻO mākou kāu koho maikaʻi loa.
Nā Papa kuhikuhi:Mea Manaʻo Hui Pū ʻIa RNA Isolation Kit Plus

Loaʻa iā mākou i kēia manawa kekahi o nā mīkini hana kiʻekiʻe loa, nā ʻenehana loea a me nā limahana, manaʻo ʻia nā ʻōnaehana hoʻokele maikaʻi a me kahi pūʻulu kūʻai akamai aloha mua / ma hope o ke kūʻai ʻana i ke kumu kūʻai no ka Rapid and High Yeild Virus Total Rna Isolation Extraction Purification Kit Nucleic Acid Detection Kit, "Making the Products of Large Quality" ʻo ia ke kumu mau loa o kā mākou ʻoihana.Hana mākou i ka hoʻoikaika mau ʻana e ʻike i ka pahuhopu o "We Will Always Hold in Pace along with the Time".
Kumukuai i oleloia noChina Universal Rna DNA Reagents a me Automatic Nucleic Acid Purification, I mea e hoʻokō ai i kā mākou pahuhopu o "ka mea kūʻai mua a me ka pōmaikaʻi like" i ka hui pū ʻana, hoʻokumu mākou i kahi hui ʻenekinia ʻoihana a me kahi hui kūʻai aku e hoʻolako i ka lawelawe maikaʻi loa e hoʻokō i nā koi o kā mākou mea kūʻai.Welina mai iā ʻoe e hui pū me mākou a hui pū me mākou.ʻO mākou kāu koho maikaʻi loa.

Hoʻopili ʻia ke kolamu

Ma hope o ka hoʻopili ʻana o ke kolamu, hoʻemi ʻia ka hua RNA a i ʻole hiki ke hoʻomaʻemaʻe i ka RNA, a haʻahaʻa ka nui o ka RNA i loaʻa.

ʻIkepili kumu maʻamau:

1. ʻAʻole pololei nā haʻawina laʻana.

ʻAʻole hiki i ka haʻihaʻi laʻana ke hoʻopaʻa ʻia ka DNA-CLEANING COLUMN, ʻoiai e pili ana i ka hua RNA a me ka maikaʻi.Paipai mākou i ka hana wili wikiwiki i loko o ka wai naikokene lawa i ka wā i haki ai ʻoe i nā laʻana, E hoʻāʻo e ʻoki i ka pā hale hāpana, membrane cell a me nā mea ʻē aʻe.No nā laʻana mea kanu o polyol polysaccharides, paipai mākou iā ʻoe e hoʻohana i ka Plant Total RNA ISOLATION KIT PLUS.

2. Ke hoʻomoʻa ʻia ka laʻana supernatant i hoʻokaʻawale ʻia me ke kolamu hoʻomaʻemaʻe DNA, hiki ke hoʻomoʻa ʻia ka wai i ʻoki ʻia.

ʻO nā sediments ʻāpana pūlima i lawe ʻia e hoʻopau ʻia ke kolamu RNA-ONLY i ka wā e hana ʻia ai ka hana adsorption RNA (e nānā i ka pae 6).Paipai mākou iā ʻoe me ke akahele i ka wā e hoʻomohu ai i kēia supernatant e pale aku i nā ʻōpala o ke kelepona i ka omo ʻana.

3. He nui loa ka helu mua.

ʻO ka hoʻohana nui ʻana i ka laʻana e hopena i ka ʻāpana hāpana piha ʻole a i ʻole ka pau ʻole o ka cell lysis e Buffer PSL1, ka hopena o ka paʻa ʻana o ke kolamu hoʻomaʻemaʻe i ka wā hoʻomaʻemaʻe.He 50 mg ka mea kanu Total RNA Isolation Kit.No nā laʻana mea kanu o polyol polysaccharides, paipai mākou iā ʻoe e hoʻāʻo iā Plant Total RNA ISOLATION KIT PLUS.

4. He haʻahaʻa loa ka mahana o ka centrifuge.

Lawe ʻia ke kaʻina hoʻokaʻawale a me ka hoʻomaʻemaʻe RNA holoʻokoʻa ma ka lumi wela (20-25°C), koe naʻe ka haʻihaʻi ʻana o ka ʻiʻo hāpana e ka wai hau.℃, hiki ke hoʻopaʻa ʻia ke kolamu hoʻomaʻemaʻe DNA a/a i ʻole kolamu RNA wale nō.Inā hiki kēia, e hoʻonoho i ka mahana centrifuge i 20-25℃, ae hoʻomaʻamaʻa mua i ka hui ʻana o ka lysis a/a i ʻole ka supernatant i hoʻohui ʻia ethanol i 37°C.

ʻAʻohe RNA i unuhi ʻia a haʻahaʻa paha ka hua RNA

Nui nā kumu e pili ana i ka pono o ka hoʻihoʻi ʻana, e like me: sample RNA content, operation method, the elution volume, etc.

Ka nānā 'ana i nā kumu ma'amau e like me lalo:

1. Ua hana ʻia kahi ʻauʻau hau a i ʻole ka wela haʻahaʻa (4 ° C) centrifugation i ka wā o ka hana.

Manaʻo: E hana ma ka lumi wela (15-25°C) i ke kaʻina holoʻokoʻa, mai hana i ka hau hau a me ka haʻahaʻa wela centrifugation.

2. Ua hoʻohaʻahaʻa ʻia ka RNA ma muli o ka mālama pono ʻole ʻana o ka hāpana a i ʻole ka mālama lōʻihi o ka hāpana.

Manaʻo Manaʻo: Pono e hoʻopaʻa koke ʻia nā laʻana i hōʻiliʻili hou ʻia i loko o ka hau hau, a laila mālama ʻia ma -80 ° C no ka manawa lōʻihi, e pale i ka hoʻoheheʻe pinepine ʻana a me ka hoʻoheheʻe ʻana o nā laʻana;a i ʻole e hoʻoinu koke i nā laʻana i loko o ka solution RNA stabilizer RNAlater solution (nā laʻana holoholona).

3.Insufficient hāpana fragmentation a me lysis alakai i blockage o ke kolamu hoomaemae.

Manaʻo Manaʻo: I ka wā e wili ai i ka ʻiʻo, e ʻoluʻolu e hoʻopaʻa pono i ka lepo, a e hoʻoneʻe koke iā ia i ka Buffer PSL1 i hoʻomākaukau mua ʻia (e hōʻoia ua hoʻohui ʻia ka māhele kūpono o β-ME, e ʻike i ka pae 1 o ke kaʻina hana).

4. Ua hoʻohui hewa ʻia ka eluent.

Manaʻo Manaʻo: E hōʻoia i ka kulu ʻana o RNase-Free ddH2O i waenakonu o ka membrane kolamu hoʻomaʻemaʻe.

5. ʻAʻole i hoʻohui ʻia ka leo kūpono o ka ethanol piha i ka Buffer PSL2 a i ʻole Buffer PRW2.

Manaʻo Manaʻo: E ʻoluʻolu e hahai i nā ʻōlelo kuhikuhi, e hoʻohui i ka leo kūpono o ka ethanol piha i ka Buffer PSL2 a me Buffer PRW2 a hui maikaʻi ma mua o ka hoʻohana ʻia ʻana o ka pahu.

6. ʻAʻole kūpono ka nui o nā kiko kiko.

Manaʻo Manaʻo: E hoʻohana i 50 mg o ka ʻiʻo no 500 μl o Buffer PSL1.ʻO ka hoʻohana ʻana i ka ʻiʻo nui e hōʻemi i ka nui o ka RNA i unuhi ʻia a e hōʻemi pū ʻia ka maʻemaʻe o ka RNA hopena.Manaʻo ikaika mākou ʻaʻole pono e ʻoi aku ka nui o ka hoʻohālike mua ma mua o 50 mg no ka hana unuhi RNA.

7.Inappropriate elution volume a incomplete elution.

Manaʻo: ʻO ka leo eluent o ke kolamu hoʻomaʻemaʻe he 50-200 μl;inā ʻaʻole ʻoluʻolu ka hopena elution, pono e hoʻolōʻihi i ka manawa ma ka lumi wela ma hope o ka hoʻohui ʻana i ka RNase-Free ddH2O preheated, e like me 5-10min.

8. He koena ethanol ka kolamu hoʻomaʻemaʻe ma hope o ka holoi ʻana me BufferPRW2.

Manaʻo Manaʻo: Inā centrifuged ka paipu ʻole no 1 min a aia nō ka ethanol i koe ma hope o ka holoi ʻana i Buffer PRW2, hiki iā ʻoe ke hoʻonui i ka manawa o ka centrifugation paipu ʻole i 2 min, a i ʻole e kau i ke kolamu hoʻomaʻemaʻe ma ka lumi wela no 5 min e wehe loa i ke koena ethanol.

9. Ua hoʻohana hewa ʻia ka pahu.

Manaʻo Manaʻo: No nā laʻana mea kanu o polyphenolic polysaccharides, me ka hoʻohana ʻana i nā pahu maʻamau e like me Plant Total RNA Isolation Kit ʻaʻole hiki ke loaʻa nā laʻana RNA kūpono.Paipai mākou iā ʻoe e hoʻohana i ka Plant Total RNA IsolationKit Plus, i hoʻolālā kūikawā ʻia no nā laʻana mea kanu polyphenolic polysaccharide.He kit i hoʻomohala kūikawā no ka unuhi ʻana i ka RNA mai nā laʻana mea kanu polyphenol a me polysaccharide.

He haʻahaʻa ka waiwai OD260/OD280

ʻO ka elution RNA me ddH2O a hoʻohana ʻia no ka heluhelu ʻana i ka spectrophotometer i loaʻa nā waiwai haʻahaʻa OD260/OD280.Manaʻo mākou e hoʻohana i ka 10 mM Tris-HCl, pH 7.5 (ma mua o ka RNase-Free ddH2O e hoʻoheheʻe i ka RNA) no ka loaʻa ʻana o nā koina OD260/OD280 pololei, e ʻike i ka "RNA Concentration and Purification Assays" ma ka ʻaoʻao 19.

Hoʻohaʻahaʻa ʻia ka RNA i hoʻomaʻemaʻe ʻia

ʻO ka maikaʻi o ka RNA i hoʻomaʻemaʻe ʻia e pili ana i nā mea e like me ka mālama ʻana i ka laʻana, ka haumia RNase, a me ka manipulation.

Ka nānā 'ana i nā kumu ma'amau:

1.ʻAʻole i mālamaʻia nā'āpana kiko i ka manawa ma hope o ka hōʻiliʻiliʻana.

Manaʻo Manaʻo: Inā ʻaʻole hoʻohana ʻia nā laʻana kikoʻī i ka manawa ma hope o ka hōʻiliʻili ʻana, e ʻoluʻolu e mālama iā lākou i loko o ka nitrogen wai i ka haʻahaʻa haʻahaʻa koke a i ʻole e hoʻololi iā lākou i -80°C no ka mālama lōʻihi ma hope o ka hoʻomaloʻo wikiwiki ʻana i loko o ka nitrogen wai, a i ʻole e hoʻokomo koke i nā laʻana i loko o ka RNA stabilizer RNAlater solution (nā mea hoʻohālike holoholona).No ka unuhi RNA, e ho'āʻo e hoʻohana i nā laʻana kikokikona i hōʻiliʻili hou ʻia.

2. Hoʻohou hou i ka maloʻo a me ka hoʻoheheʻe ʻana i nā ʻāpana kiko.

Manaʻo Manaʻo: I ka mālama ʻana i nā laʻana ʻiʻo, ʻoi aku ka maikaʻi e ʻoki i nā ʻāpana liʻiliʻi no ka mālama ʻana, a lawe i kahi ʻāpana o ia mau mea i ka wā e hoʻohana ai i mea e pale ai i ka hōʻino ʻana o ka RNA ma muli o ka hoʻomaloʻo pinepine ʻana a me ka hoʻoheheʻe ʻana o nā laʻana.

3. Hoʻokomo ʻia ʻo RNase i loko o ka lumi hana a ʻaʻole i hoʻohana ʻia i nā mīkina lima hoʻolele, masks, etc.

Manaʻo Manaʻo: ʻoi aku ka maikaʻi o nā hoʻokolohua RNA extraction i nā hana RNA kaʻawale, a pono e hoʻomaʻemaʻe ʻia ka papa ʻaina ma mua o ka hoʻokolohua, a pono e ʻaʻahu ʻia nā mīkina lima a me nā masks i ka wā o ka hoʻokolohua e pale aku ai i ka hōʻino ʻana o RNA ma muli o ka hoʻokomo ʻana o RNase i ka nui loa.

4. Ua haumia ka reagent e RNase i ka wā e hoʻohana ai.

Manaʻo Manaʻo: E hoʻololi me kahi pūʻulu hou o nā pahu hōʻiliʻili RNA piha no nā hoʻokolohua pili.

5. ʻO nā paipu centrifuge a me nā ʻōlelo aʻoaʻo pipette i hoʻohana ʻia no ka hoʻoponopono RNA ua haumia me RNase.

Manaʻo Manaʻo: E hōʻoia i ka RNase-Free nā paipu centrifuge, nā ʻōlelo aʻoaʻo pipette, nā pipette, a me nā mea ʻē aʻe i hoʻohana ʻia i ka unuhi RNA.

Nā Papa kuhikuhi:

Mea Manaʻo Hui Pū ʻIa RNA Isolation Kit Plus