Ngwakọta mkpokọta RNA kewapụ ihe ọkụkụ gbakwunyere Ngwakọta RNA Purificaiton maka ihe ọkụkụ bara ụba na Polysaccharides na Polyphenols.
Nkọwa ngwa:
Cat.No.RE-05021/05022/05024
Maka ịdị ọcha nke mkpokọta RNA site na nlele osisi n'ozuzu nwere nnukwu polysaccharide na ihe polyphenol.
Wepụ mkpokọta RNA dị elu ngwa ngwa site na nlele osisi nwere ọdịnaya dị elu nke polysaccharides na polyphenols.
RNase-efu n'iji kọlụm na-ehicha DNA
Mfe-arụ ọrụ niile na-arụcha na ụlọ okpomọkụ
Ngwa ngwa-enwere ike ịrụcha ọrụ n'ime nkeji iri atọ
Safe — ọ dịghị organic reagent eji 
Nkọwapụta
Nkwadebe 50, 200 Preps
Ngwa ahụ na-eji kọlụm ogho na usoro nke Foregene mepụtara, nke nwere ike wepụ RNA zuru oke na ịdị ọcha dị elu site na anụ ahụ osisi dị iche iche nwere nnukwu polysaccharides ma ọ bụ polyphenols.Ọ na-enye kọlụm DNA- Cleaning nke nwere ike wepụ DNA genomic ngwa ngwa site na lysate na 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 |
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.
■ Kwesịrị ekwesị maka ịsacha RNA site na ihe atụ osisi nke polysaccharides na polyphenols.
∎ 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 ngwaahịa
∎ Ngwa mgbada ala: njikọ cDNA nke mbụ, RT-PCR, cloning molecular, Northern Blot, wdg.
■ Ihe nlele: anụ ahụ osisi dị ọhụrụ ma ọ bụ oyi kpọnwụrụ nke polysaccharides na polyphenols
■ Usoro onunu ogwu: 50mg osisi anụ ahụ
■ Ike njide RNA kacha nke kọlụm nchacha: 80 μg
■ Olu olu: 50-200 μl
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 ahịhịa ọhụrụ) nwere nnukwu 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 Shelf ndụ
Enwere ike ịchekwa ngwa a maka ọnwa 24 n'okpuru ọnọdụ akọrọ na ụlọ okpomọkụ (15-25 ℃);Ọ bụrụ na ọ dị mkpa ka ịchekwa ya ogologo oge, enwere ike ịchekwa ya na 2-8 ℃.
Enwere ike itinye ihe nchekwa PSL1 na 4 ℃ maka ọnwa 1 mgbe agbakwunyere β-mercaptoethanol (a na-atụ aro ka ịgbakwunye ya n'otu oge nnwale).
Akwụnyere kọlụm
Ka etinyechara kọlụm ahụ, mkpụrụ RNA na-ebelata ma ọ bụ ọ gaghị ekwe omume ime ka RNA dị ọcha, yana oke RNA enwetara dị ala.
Nyocha ihe kpatara ya:
1. Ntụle nkwụsịtụ adịghị nke ọma.
Njiji ihe atụ anaghị eme ka egbochie kọlụm DNA-CLEANING kpamkpam, ebe ọ na-emetụta mkpụrụ RNA na ịdị mma.Anyị na-akwado ngwa ngwa-egweri ọrụ na a zuru ezu mmiri mmiri nitrogen mgbe ị mebiri samples, Gbalịa na-echipịa sample cell mgbidi, cell akpụkpọ ahụ na ndị ọzọ na anụ ahụ.Maka nlele osisi nke polyol polysaccharides, anyị na-akwado ka ị jiri Plant Total RNA ISOLATION KIT PLUS.
2. Mgbe ejiri DNA-Cleaning Kọlụm na-amịpụta ihe nlere nke kewapụrụ ekewa, enwere ike ikuru mmiri na-agbajisị cell.
Sedimentị gbajiri sel nke ewepụrụ ga-ebute kọlụm RNA-ONLY nke a ga-egbochi mgbe arụ ọrụ mgbasa ozi RNA na-arụ (lee nzọụkwụ 6).Anyị na-akwado gị nke ọma mgbe ị na-amịpụta ihe a na-ahụ anya iji zere irighiri irighiri cell.
3. Sample mbụ ego dị ukwuu.
Ojiji nke oke nlere ga-ebute nkewa nlere ezughị ezu ma ọ bụ lysis cell ezughị ezu site na Buffer PSL1, na-ebute mgbochi nke kọlụm nchacha n'oge a na-asachapụ.Ngwakọta mkpokọta RNA kewapụrụ ihe ọkụkụ nke ọ bụla n'otu n'otu arụrụ arụ ọrụ bụ 50 mg.Maka ihe atụ osisi nke polyol polysaccharides, anyị na-akwado ka ị nwaa Plant Total RNA ISOLATION KIT PLUS.
4. Okpomọkụ nke centrifuge dị oke ala.
A na-eme ikewapụ RNA niile na usoro ịsacha na ụlọ okpomọkụ (20-25°C), ma e wezụga na mmiri mmiri nitrogen agbajiwo anụ ahụ sample. Okpomọkụ nke ụfọdụ centrifuges cryogenic dị ala karịa 20℃, nke nwere ike ibute ngọngọ nke kọlụm DNA-Cleaning na/ma ọ bụ naanị kọlụm RNA.Ọ bụrụ na nke a emee, debe okpomọkụ centrifuge na 20-25℃, najide n'aka na ngwakọta lysis na/ma ọ bụ ethanol gbakwunyere na-ekpo ọkụ na-ekpo ọkụ na 37°C.
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. A na-emebi RNA ahụ n'ihi nchekwa na-ezighị 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.Enweghị oke nlele nkewa 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 ddH2O na-enweghị RNase 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.Na-ekwesịghị ekwesị elution olu ma ọ bụ ezughị ezu elution.
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 n'ime ụlọ okpomọkụ mgbe agbakwunyere ddH2O na-enweghị ọkụ RNase, dị ka 5-10min.
8.The kọlụm dị ọcha nwere ethanol fọdụrụnụ mgbe ịsacha na BufferPRW2.
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 elution na ddH2O ma jiri 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 iji elute RNA) iji nweta ụkpụrụ OD260/OD280 ziri ezi, lee "Ntụle 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:
Ngwakọta mkpokọta RNA kewapụrụ ihe ọkụkụ gbakwunyere ntuziaka ntuziaka
