Ngwakọta mkpokọta RNA kewapụ ihe ọkụkụ mkpokọta RNA Purificaiton Kit maka Osisi dị ala na Polysaccharides na Polyphenols.
Nkọwapụta
Nkwadebe 50, 200 Preps
Ngwa ahụ na-eji kọlụm ogho na usoro nke Foregene mepụtara, nke nwere ike wepụta RNA dị ọcha na ịdị elu nke ọma site na anụ ahụ osisi dị iche iche nwere polysaccharides dị ala na ọdịnaya polyphenols.Maka nlele osisi nwere nnukwu polysaccharides ma ọ bụ polyphenols, a na-atụ aro ka iji Plant Total RNA Isolation Plus Kit nweta nsonaazụ mmịpụta RNA ka mma.Ngwa ahụ na-enye kọlụm DNA- Cleaning nke nwere ike wepu DNA genomic n'ụzọ dị mfe na lysate nke anụ ahụ.Kọlụm RNA naanị nwere ike jikọta RNA nke ọma.Ngwa ahụ nwere ike ịhazi ọnụ ọgụgụ dị ukwuu nke ihe nlele n'otu oge.
Sistemu niile enweghị RNase, yabụ na RNA emechara adịghị eme ka ọ dị ala.Buffer PRW1 na Buffer PRW2 nwere ike hụ na RNA enwetaghị protein, DNA, ion, na ogige organic emerụghị ya.
Ngwa ngwa
| Buffer PSL1, ihe nchekwa PS, ihe nchekwa PSL2 |
| Ihe nchekwa PRW1, ihe nchekwa PRW2 |
| ddH na-efu RNase2O, kọlụm na-ehicha DNA |
| RNA-naanị kọlụm |
| Ntuziaka |
Atụmatụ & uru
■ Arụ ọrụ na ụlọ okpomọkụ (15-25 ℃) n'ime usoro dum, na-enweghị ice bat na obere okpomọkụ centrifugation.
∎ Ngwa RNase zuru oke, enweghị mkpa ichegbu onwe gị maka mmebi RNA.
∎ kọlụm DNA- Cleaning na-ejikọta kpọmkwem na DNA, ka ngwa ahụ wee nwee ike wepụ nje DNA nje na-agbakwunyeghị DNA.
∎ Mkpụrụ RNA dị elu: Kọlụm naanị RNA nwere ike sachaa RNA nke ọma.
∎ Ọsọ ọsọ: dị mfe iji rụọ ọrụ, enwere ike mezue ya n'ime nkeji iri atọ.
■ Nchekwa: ọ dịghị organic reagenti a chọrọ.
■ Ọdịmma dị elu: Iberibe RNA ndị a sachara dị ọcha, enweghị protein na adịghị ọcha ndị ọzọ, ma nwee ike izute ngwa nnwale dị iche iche nke ala.
Ngwa ngwa
Ọ dabara maka mmịpụta na ịdị ọcha nke mkpokọta RNA site na nlele anụ ahụ dị ọhụrụ ma ọ bụ oyi kpọnwụrụ (karịsịa anụ ahụ akwụkwọ osisi ọhụrụ) nwere obere polysaccharide na ọdịnaya polyphenol.
Usoro ọrụ
Eserese
Plant Total RNA Isolation Kit Plus na-ahazi 50mg nke akwụkwọ ọhụrụ nke polysaccharides na polyphenols, na 5% RNA dị ọcha ka a nwalere site na electrophoresis.
1: banana
2: Ginkgo
3: owu
4: pọmigranet
Nchekwa na ndụ nchekwa
Enwere ike ịchekwa ngwa ahụ maka ọnwa 12 n'ime ụlọ okpomọkụ (15-25 ℃) na ebe akọrọ, yana 2-8 ℃ maka ogologo oge (ọnwa 24).
Enwere ike ịchekwa ihe nchekwa PSL1 na 4 ℃ maka ọnwa 1 mgbe ịgbakwunye 2-hydroxy-1-ethanethiol (nhọrọ).
Ntuziaka nyocha nsogbu
Nyocha na-esonụ nke nsogbu ị nwere ike izuteNgụkọta ihe ọkụkụRNA extraction will help you with your experiments. In addition, for other experimental or technical problems in addition to operating instructions and problem analysis, we have dedicated technical support to help you. If you have any needs, please contact us at: 028-83360257 or E-mali : Tech@foregene.com.
Akpọchiela kọlụm atụ ogho
Mgbochi nke kọlụm ntugharị ga-eme ka mkpụrụ RNA belata ma ọ bụ ọbụlagodi enweghị ike ịsacha iji nweta RNA, ogo RNA enwetara ga-adị ala.
Nyocha ihe kpatara ya:
1. A naghị agbaji ihe nlele ahụ kpamkpam.
Nkewa nlere ezughi oke nwere ike igbochi kọlụm DNA-Cleaning, nke nwekwara ike imetụta mkpụrụ RNA na ịdịmma.Anyị na-akwado na mgbe ị na-eme nkewa sample, ngwa ngwa gwerie mmiri mmiri nitrogen zuru oke iji mebie anụ ahụ dị ka mgbidi cell na akpụkpọ anụ cell nke ihe nlele ahụ dị ka o kwere mee.Maka nlele osisi nke polyphenol polysaccharides, anyị na-akwado ka ị jiri Plant Total RNA Isolation Kit Plus.
2.Aspirate kọlụm DNA- Cleaning dịpụrụ adịpụ supernatant, chọọ pellet irighiri mkpụrụ ndụ enwere ike.
Pellet irighiri mkpụrụ ndụ aspirated nwere ike mechie kọlụm RNA naanị n'oge usoro mgbasa ozi RNA (lee nzọụkwụ 5 nke usoro, nzọụkwụ 6 nke usoro polysaccharide polyphenol).Anyị na-akwado na a ga-akpachapụ anya mgbe ị na-achọ ihe karịrị nke a iji zere irighiri ihe ndị dị na cell.
3. Ọnụ ego mbụ nke nlele dị oke.
Ojiji nke oke nlere ga-ebute nkewa nlere ezughi oke ma ọ bụ lysis nke mkpụrụ ndụ ezughị ezu site na Buffer PRL1 ma ọ bụ Buffer PSL1, na-ebute kọlụm nchacha ọcha maka arụ ọrụ dị ọcha.Plant Total RNA Isolation Kit nwere mbido mbụ nke 50 mg kwa otu nchacha nke ihe nlele eji arụ ọrụ.Maka ihe atụ osisi nke polyphenol polysaccharides, anyị na-akwado ka ị nwaa Plant Total RNA Isolation Kit Plus.
4. Okpomọkụ nke centrifuge dị oke ala.
Mwepu RNA na ịdị ọcha belụsọ n'ihi mmebi nitrogen mmiri mmiri nke anụ ahụ nlele, a na-eme usoro niile na ụlọ okpomọkụ (20-25 Celsius C).Ụfọdụ centrifuges na-ekpo ọkụ nwere okpomọkụ n'okpuru 20 Celsius C, nke nwere ike ịkpata mgbochi na kọlụm DNA-Cleaning na/ma ọ bụ naanị kọlụm RNA.Ọ bụrụ na nke a emee, tọọ okpomọkụ centrifuge ka ọ bụrụ 20-25 Celsius C wee kpoo ngwakọta lysis na/ma ọ bụ mgbakwunye nke nkewa nke ethanol na 37 Celsius.
Ọ nweghị RNA ewepụtara ma ọ bụ mkpụrụ RNA dị ala
A na-enwekarị ọtụtụ ihe na-emetụta arụmọrụ mgbake, dị ka: sample ọdịnaya RNA, usoro ọrụ, elution olu, wdg.
Nyocha nke ihe na-akpatakarị dị ka n'okpuru:
1.A na-eme centrifugation ice ma ọ bụ obere okpomọkụ (4 ° C) n'oge ọrụ ahụ.
Aro: Na-arụ ọrụ na ụlọ okpomọkụ (15-25 ° C) na usoro dum, emela ice bat na obere okpomọkụ centrifugation.
2.Emebiri RNA ahụ n'ihi nchekwa na-ezighi ezi nke ihe nlele ma ọ bụ nchekwa ogologo oge nke ihe nlele ahụ.
Nkwanye: A ga-ekpochapụ ihe nlele ndị anakọtara ọhụrụ na nitrogen mmiri mmiri ngwa ngwa, wee chekwaa ya na -80 Celsius ruo ogologo oge, zere ịza oyi ugboro ugboro na thawing samples;ma ọ bụ gbanye ihe nlele ozugbo na ngwọta RNA stabilizer RNAlater (ihe nlele anụmanụ).
3.Enweghi ike nkewa sample na lysis na-eduga na ngọngọ nke kọlụm dị ọcha.
Aro: Mgbe ị na-egweri anụ ahụ, biko hụ na anụ ahụ agbadala nke ọma, ma bufee ya ngwa ngwa na Buffer PSL1 akwadoro (kwenye na agbakwunyere oke β-ME ziri ezi, lee nzọụkwụ 1 nke usoro).
4.Agbakwunyere eluent na-ezighi ezi.
Aro: Gbaa mbọ hụ na ddH enweghị RNase2O na-adaba n'etiti akpụkpọ ahụ kọlụm dị ọcha.
5.Etinyeghị olu ziri ezi nke ethanol zuru oke na Buffer PSL2 ma ọ bụ Buffer PRW2.
Aro: Biko soro ntuziaka a, tinye nha ziri ezi nke ethanol zuru oke na Buffer PSL2 na Buffer PRW2 wee gwakọta nke ọma tupu ejiri ngwa ahụ.
6. Ọnụ ọgụgụ nke anụ ahụ adịghị mma.
Aro: Jiri 50 mg nke anụ ahụ kwa 500 μl nke Buffer PSL1.Iji anụ ahụ buru ibu ga-ebelata ọnụọgụ RNA ewepụtara yana ịdị ọcha nke RNA ga-esi na ya pụta ga-ebelatakwa.Anyị na-akwado ike na usoro nlele mbụ ekwesịghị gafere 50 mg kwa ọrụ mmịpụta RNA.
7. Mpịakọta elution na-ekwesịghị ekwesị ma ọ bụ elution ezughị ezu.
Aro: Oke eluent nke kọlụm dị ọcha bụ 50-200 μl;Ọ bụrụ na mmetụta elution adịghị afọ ojuju, a na-atụ aro ịgbatị oge na okpomọkụ ụlọ mgbe agbakwunyere ddH na-enweghị ọkụ RNase.2O, dị ka 5-10min.
8.The kọlụm dị ọcha nwere ethanol fọdụrụnụ mgbe ịsacha na Buffer PRW2.
Aro: Ọ bụrụ na centrifuged tube efu maka 1 min ma a ka nwere ethanol fọdụrụ mgbe ịsachara na Buffer PRW2, ị nwere ike ịbawanye oge nke tube centrifugation efu na 2 min, ma ọ bụ tinye kọlụm dị ọcha na ụlọ okpomọkụ maka 5 min iji wepụ ethanol fọdụrụnụ kpamkpam.
9.A na-eji ngwa ahụ eme ihe na-ezighi ezi.
Aro: Maka ihe atụ osisi nke polysaccharides polyphenolic, iji ngwa ndị a na-ahụkarị dịka Plant Total RNA Isolation Kit nwere ike ọ gaghị enwe ike nweta ụdị RNA dị mma.Anyị na-akwado ka ị jiri Plant Total RNA IsolationKit Plus, nke emebere nke ọma maka nlele osisi polyphenolic polysaccharide.Ngwa emepụtara nke ọma maka ịwepụta RNA na ihe atụ osisi polyphenol na polysaccharide.
OD260/OD280 uru dị ala
RNA elu na ddH2O na ejiri ya mee ihe maka ọgụgụ spectrophotometer na-arụpụta ụkpụrụ OD260/OD280 dị ala.Anyị na-akwado iji 10 mM Tris-HCl, pH 7.5 (kama RNase-Free ddH2O to elute RNA) iji nweta ụkpụrụ OD260/OD280 ziri ezi, lee “ RNA Concentration and Purification Assays” na ibe 19.
A na-emebi RNA dị ọcha
Ọdịmma RNA dị ọcha metụtara ihe ndị dị ka ichekwa nlele, mmetọ RNase, na nhịahụ.
Nyocha nke ihe na-akpatakarị:
A naghị echekwa ihe nlele 1.Tissue na oge mgbe nchịkọta.
Nkwanye: Ọ bụrụ na ejighị ihe nlele anụ ahụ n'oge nchịkọta, biko chekwaa ha na mmiri mmiri nitrogen na obere okpomọkụ ozugbo ma ọ bụ bufee ya na -80 Celsius maka nchekwa ogologo oge mgbe oyi na-ekpo ọkụ ngwa ngwa na nitrogen mmiri mmiri, ma ọ bụ gbanye ihe nlele ozugbo na ngwọta RNA stabilizer RNAlater (ntụle anụmanụ).Maka mmịpụta RNA, gbalịa iji nlele anụ ahụ anakọtara ọhụrụ.
2.Na-emegharị ugboro ugboro na thawing nke anụ ahụ n'omume.
Aro: Mgbe ị na-echekwa ihe nlele anụ ahụ, ọ kacha mma ịkpụ ha n'ime obere iberibe maka nchekwa, wee wepụta akụkụ ha mgbe ị na-eji ha iji zere mmebi nke RNA nke oyi na-eme ugboro ugboro na agbaze nke samples.
A na-ewebata 3.RNase n'ime ụlọ ọrụ ma ọ bụ na-eyighị uwe, mkpuchi, wdg.
Aro: A na-eme nnwale mmịpụta RNA kacha mma na arụ ọrụ RNA dị iche iche, a ga-ehichapụkwa tebụl ụlọ nyocha tupu nnwale ahụ, a ga-eyiri uwe aka na ihe mkpuchi nwere ike iyi n'oge nnwale ahụ iji zere mbibi RNA kpatara site na iwebata RNase ruo oke.
4.The reagent na-emerụ site RNase n'oge eji.
Aro: Dochie na usoro ọhụrụ nke ngwa mkpokọta RNA ihe ọkụkụ maka nnwale ndị metụtara ya.
5.The centrifuge tubes na pipette ndụmọdụ eji eme ihe RNA na-emetọ ya na RNase.
Aro: Gbaa mbọ hụ na tubes centrifuge, ndụmọdụ pipette, pipettes, wdg. ejiri na mmịpụta RNA bụ RNase-efu.
Akwụkwọ ntuziaka:
