ʻO kā mākou mau lako lako pono a me ka mana maikaʻi maikaʻi maikaʻi loa ma nā pae āpau o ka hana ʻana e hiki ai iā mākou ke hōʻoia i ka piha o ka mea kūʻai aku i ka hale kūʻai pololei ʻo China Techstar Nest Mgpure Nucleic Acid Purification Kit Extraction Kit Rna Isolation Kit, "E hana i nā huahana a me nā hopena o ka Large Quality" ʻo ia ka pahuhopu mau loa o kā mākou hui.Hana mākou i nā hoʻoikaika ʻole e ʻike i ka manaʻo o "E hoʻomau mau mākou i ka wikiwiki me nā manawa āpau".
ʻO kā mākou mau lako lako pono a me ka mana maikaʻi maikaʻi maikaʻi loa ma nā pae āpau o ka hana ʻana e hiki ai iā mākou ke hōʻoia i ka hauʻoli o ka mea kūʻai aku.Kina Nucleic Acid Reagent, ʻO ka pahu hoʻoheheʻe DNA/Rna, Hāʻawi mākou i ka lawelawe kūpono, pane wikiwiki, hāʻawi manawa kūpono, maikaʻi maikaʻi a me ke kumukūʻai maikaʻi loa i kā mākou mea kūʻai.ʻO ka ʻoluʻolu a me ka hōʻaiʻē maikaʻi i kēlā me kēia mea kūʻai aku kā mākou mea nui.Kūleʻa mākou i kēlā me kēia kikoʻī o ka hoʻoponopono kauoha no nā mea kūʻai aku a hiki i ka loaʻa ʻana o nā huahana palekana a maikaʻi me ka lawelawe logistics maikaʻi a me ke kumukūʻai waiwai.Ma muli o kēia, kūʻai maikaʻi ʻia kā mākou huahana ma nā ʻāina ma ʻApelika, ka Mid-East a me Asia Hema.

Nā kikoʻī

50 Preps, 200 Preps Ke hoʻohana nei ka pahu i ke kolamu spin a me ka formula i hoʻomohala ʻia e Foregene, hiki iā ia ke hoʻokaʻawale maikaʻi i ka RNA piha maʻemaʻe kiʻekiʻe a kiʻekiʻe mai nā ʻiʻo mea kanu me nā polysaccharides kiʻekiʻe a i ʻole polyphenols.Hāʻawi ia i ke kolamu hoʻomaʻemaʻe DNA e hiki ke wehe maʻalahi i ka DNA genomic mai ka supernatant a me ka lysate kiko.Hiki i ke kolamu RNA-wale ke hoopaa pono i ka RNA.Hiki i ke kit ke hana i ka nui o nā laʻana i ka manawa like.

ʻAʻole i loko o ka ʻōnaehana holoʻokoʻa RNase, no laila ʻaʻole e hoʻohaʻahaʻa ʻia ka RNA i hoʻomaʻemaʻe ʻia.Hiki i ka Buffer PRW1 a me ka Buffer PRW2 ke hōʻoia ʻaʻole i haumia ka RNA i loaʻa e ka protein, DNA, nā ion, a me nā pūhui organik.

Nā ʻāpana pahu

Nā hiʻohiʻona & pono

■ Ka hana ma ka lumi wela (15-25 ℃) ma ke kaʻina holoʻokoʻa, me ka ʻauʻau hau a me ka centrifugation haʻahaʻa wela.■ Hoʻopiha piha RNase-Free, ʻaʻohe pono e hopohopo e pili ana i ka hōʻino ʻana o RNA.■ He kūpono no ka hoʻomaʻemaʻe ʻana i ka RNA mai nā laʻana mea kanu o polysaccharides a me polyphenols.■ Hoʻopili pono ʻia ke kolamu hoʻomaʻemaʻe DNA i ka DNA, i hiki ai i ka pahu ke hoʻoneʻe i ka ʻino DNA genomic me ka hoʻohui ʻole ʻana i ka DNase.■ Loaʻa RNA kiʻekiʻe: Hiki i ke kolamu RNA-wale a me ka hoʻohālikelike kū hoʻokahi ke hoʻomaʻemaʻe pono i ka RNA.■ ʻO ka wikiwiki wikiwiki: maʻalahi e hana a hiki ke hoʻopau i loko o 30 mau minuke.■ Palekana: ʻaʻole koi ʻia ka reagent organik.■ Kiʻekiʻe: ʻO nā ʻāpana RNA i hoʻomaʻemaʻe ʻia he maʻemaʻe kiʻekiʻe, ʻaʻohe protein a me nā mea haumia ʻē aʻe, a hiki ke hālāwai me nā noi hoʻokolohua like ʻole.

Nā palena huahana

■ Nā mea hoʻohana i lalo: hoʻohui mua cDNA, RT-PCR, molecular cloning, Northern Blot, etc. ■ Laʻana: ʻO nā mea kanu hou a maloʻo paha o nā polysaccharides a me nā polyphenols ■ ʻAi: 50mg mea kanu mea kanu ■ ʻO ka hiki ke hoʻopaʻa RNA kiʻekiʻe o ke kolamu hoʻomaʻemaʻe: 80 μg ■ Ka nui o ka elution: 50-200 μl

Ke noi pahu

He kūpono ia no ka unuhi ʻana a me ka hoʻomaʻemaʻe ʻana i ka RNA a pau mai nā laʻana o nā mea kanu hou a i ʻole ka hau (ʻoi aku ka ʻiʻo o ka lau mea kanu hou) me ka polysaccharide kiʻekiʻe a me ka polyphenol content.

Ka holo hana

Diagram

ʻO ka Plant Total RNA Isolation Kit Plus i hana ʻia he 50mg o nā lau hou o polysaccharides a me polyphenols, a ua hoʻāʻo ʻia ʻo 5% RNA i hoʻomaʻemaʻe ʻia e ka electrophoresis.1: Ka maiʻa 2: Ginkgo 3: Ka pulupulu 4: Pomegerane

Ka mālama ʻana a me ka mālama ʻana

Hiki ke mālama ʻia kēia kit no 24 mau mahina ma lalo o nā kūlana maloʻo ma ke ana wela (15-25 ℃);inā pono e mālama ʻia no ka manawa lōʻihi, hiki ke mālama ʻia ma 2-8 ℃.Hiki ke waiho ʻia ka Buffer PSL1 ma 4 ℃ no 1 mahina ma hope o ka hoʻohui ʻana i ka β-mercaptoethanol (manaʻo ʻia e hoʻohui iā ia i ka manawa like o ka hoʻokolohua). o kā mākou hui.Hana mākou i nā hoʻoikaika ʻole e ʻike i ka manaʻo o "E hoʻomau mau mākou i ka wikiwiki me nā manawa āpau".
Hale hana pololeiKina Nucleic Acid Reagent, ʻO ka pahu hoʻoheheʻe DNA/Rna, Hāʻawi mākou i ka lawelawe kūpono, pane wikiwiki, hāʻawi manawa kūpono, maikaʻi maikaʻi a me ke kumukūʻai maikaʻi loa i kā mākou mea kūʻai.ʻO ka ʻoluʻolu a me ka hōʻaiʻē maikaʻi i kēlā me kēia mea kūʻai aku kā mākou mea nui.Kūleʻa mākou i kēlā me kēia kikoʻī o ka hoʻoponopono kauoha no nā mea kūʻai aku a hiki i ka loaʻa ʻana o nā huahana palekana a maikaʻi me ka lawelawe logistics maikaʻi a me ke kumukūʻai waiwai.Ma muli o kēia, kūʻai maikaʻi ʻia kā mākou huahana ma nā ʻāina ma ʻApelika, ka Mid-East a me Asia Hema.

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.