Plant Yese RNA Isolation Kit Yese RNA Purificaiton Kit yeChirimwa Yakaderera muPolysaccharides uye Polyphenols
Zvinotsanangurwa
50 Preps, 200 Preps
Iyo kit inoshandisa iyo spin column uye fomula yakagadziridzwa neForegene, iyo inogona kunyatso kuburitsa yakakwira-kuchena uye yemhando yepamusoro RNA kubva kwakasiyana siyana matishu ane yakaderera polysaccharides uye polyphenols zvirimo.Kune zvidyarwa masampuli ane yakakwira polysaccharides kana polyphenols zvemukati, zvinokurudzirwa kushandisa Plant Yese RNA Isolation Plus Kit kuti uwane zvirinani RNA yekubvisa mhedzisiro.Iyo kit inopa iyo DNA-Kuchenesa koramu iyo inogona nyore kubvisa genomic DNA kubva kune supernatant uye tishu lysate.RNA-chete column inogona kunyatsosunga RNA.Iyo kit inogona kugadzirisa nhamba huru yemasampuli panguva imwe chete.
Iyo yese system haina RNase, saka iyo yakacheneswa RNA haizodzikisirwe.Buffer PRW1 uye Buffer PRW2 inogona kuvimbisa kuti RNA inowanikwa haina kusvibiswa neprotein, DNA, ions, uye organic compounds.
Kit zvikamu
| Buffer PSL1, Buffer PS, Buffer PSL2 |
| Buffer PRW1, Buffer PRW2 |
| RNase-Yemahara ddH2O, DNA-Cleaning Column |
| RNA-Chete Column |
| Mirayiridzo |
Zvimiro&zvakanakira
■ Kushanda pakamuri tembiricha (15-25 ℃) mukati mese muitiro, pasina kugeza nechando uye yakaderera tembiricha centrifugation.
■ Yakazara kit RNase-Yemahara, hapana chikonzero chekunetseka nezveRNA kudzikira.
∎ DNA-Cleaning Column inosungirira kuDNA, kuitira kuti kit ibvise kusvibiswa kweDNA yegenomic pasina kuwedzera DNase.
■ Yakakwira RNA goho: RNA-chete Column uye yakasarudzika fomula inogona kunyatsochenesa RNA.
■ Kukurumidza kumhanya: nyore kushanda uye inogona kupedzwa mukati memaminitsi makumi matatu.
■ Kuchengetedza: hapana organic reagent inodiwa.
■ Hunhu hwepamusoro: Zvimedu zveRNA zvakacheneswa ndezvekuchena kwepamusoro, hazvina mapuroteni nehumwe tsvina, uye zvinokwanisa kusangana nemhando dzakasiyana dzekuedza dzepasi.
Kit application
Yakakodzera kubviswa uye kucheneswa kweRNA yakazara kubva kutsva kana chando matishu echirimwa samples (kunyanya nyowani yemashizha emuti wetishu) ine yakaderera polysaccharide uye polyphenol zvemukati.
Kuyerera kwebasa
Mufananidzo
Plant Yese RNA Isolation Kit Plus yakagadziriswa 50mg yemashizha matsva epolysaccharides uye polyphenols, uye 5% yakacheneswa RNA yakaedzwa ne electrophoresis.
1: Banana
2: Ginkgo
3: Donje
4: Damba
Kuchengetedza uye hupenyu hwesherufu
Iyo kit inogona kuchengetwa kwemwedzi gumi nemiviri patembiricha yekamuri (15-25 ℃) munzvimbo yakaoma, uye 2-8 ℃ kwenguva yakareba (24 mwedzi).
Buffer PSL1 inogona kuchengetwa pa4 ℃ kwemwedzi 1 mushure mekuwedzera 2-hydroxy-1-ethanethiol(kusarudza).
Dambudziko rekuongorora Guide
Ongororo inotevera yezvinetso zvaungasangana nazvoPlant TotalRNA 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.
Iyo spin column yakavharwa
Kuvharika kweiyo spin column kuchaita kuti goho reRNA rideredzwe kana kutotadza kucheneswa kuti riwane RNA, uye mhando yeRNA inowanikwa ichave yakaderera.
Kuongororwa Kwechikonzero Chinozivikanwa:
1. Muenzaniso hauna kuputswa zvachose.
Kupatsanurwa kwemuenzaniso kusina kukwana kunogona kuvhara DNA-Kuchenesa Column, iyo inogona kukanganisa goho reRNA uye mhando.Isu tinokurudzira kuti kana uchiita kupatsanurwa kwemuenzaniso, kurumidza kugaya muhuwandu hwakakwana hwenitrogen yemvura kupwanya matishu akadai semadziro maseru uye maseru membranes emasampuli zvakanyanya sezvinobvira.Kune zvidyarwa zvepolyphenol polysaccharides, isu tinokurudzira kuti ushandise Plant Yese RNA Isolation Kit Plus.
2.Aspirate the DNA-Cleaning Column isolated supernatant, aspirate the possible cell debris pellet.
Aspirated cell debris pellet inogona kuvhara iyo RNA-chete Column panguva yeRNA adsorption maitiro (ona nhanho yechishanu yemaitiro, nhanho yechitanhatu yepolysaccharide polyphenol maitiro).Isu tinokurudzira kuti kungwarira kunofanirwa kutorwa kana uchishuvira iyi supernatant kudzivirira maseru marara ari kushuvira.
3. Nhamba yekutanga yemuenzaniso yakakura zvikuru.
Kunyanyisa kushandiswa kwemuenzaniso kunozokonzera kusakwana kwemuenzaniso kupatsanurwa kana kusakwana lysis yemaseru neBuffer PRL1 kana Buffer PSL1, zvichikonzera koramu yakavharika yekuchenesa yemabasa ekuchenesa.Plant Yese RNA Isolation Kit ine yekutanga yepamusoro ye50 mg pakucheneswa kumwe kwemuenzaniso wakashandiswa.Kune zvidyarwa zvepolyphenol polysaccharides, isu tinokurudzira kuti uedze iyo Yese Yese RNA Isolation Kit Plus.
4. Kupisa kwe centrifuge kwakanyanya kuderera.
Yese RNA yekuzviparadzanisa uye kucheneswa kunze kwekuvhiringidzwa kwenitrogen yemvura yemuenzaniso tishu, matanho ese anoitwa patembiricha yekamuri (20-25 ° C).Mamwe macentrifuge akadzikama ane tembiricha iri pasi pe20 °C, izvo zvinogona kukonzera kuvharika muDNA-Cleaning Column uye/kana RNA-chete Column.Kana izvi zvikaitika, isa tembiricha yecentrifuge kusvika 20-25 °C uye pre-kudziisa musanganiswa welysis uye/kana kuwedzera ethanol yekuparadzanisa supernatant kusvika 37 °C.
Hapana RNA yakabviswa kana RNA goho rakaderera
Pane kazhinji zvinhu zvakawanda zvinokanganisa kugona kudzoreredza, senge: sampuli RNA yemukati, nzira yekushanda, iyo elution vhoriyamu, nezvimwe.
Kuongororwa kwezvikonzero zvakajairika sezvinotevera:
1.Kushamba kwechando kana kutonhora kwakadzika (4 ° C) centrifugation yakaitwa panguva yekushanda.
Zano: Shanda pane tembiricha yekamuri (15-25 ° C) mukuita kwese, usaite ice bath uye yakaderera tembiricha centrifugation.
2.Iyo RNA yakaderedzwa nekuda kwekuchengetedza zvisina kunaka kwemuenzaniso kana kuchengetedzwa kwenguva refu kwemuenzaniso.
Kurudziro: Zvichangobva kuunganidzwa samples dzinofanirwa kukurumidza kuomeswa nechando mumvura ine nitrogen, yozochengetwa pa -80°C kwenguva yakareba, kudzivisa kuomesa nechando uye kunyungudutswa kwemasampuli;kana kuti nekukasira kunyudza masampuli muRNA stabilizer RNAlater mhinduro (samples dzemhuka).
3.Kusakwana kupatsanurwa kwemuenzaniso uye lysis inotungamira mukuvharika kwekoramu yekuchenesa.
Mazano: Paunenge uchikuya mutsara, tapota chengetedza kuti mutsara wakakwana pasi, uye nekukurumidza uende nayo kuBuffer pre-prepared PSL1 (simbisa kuti chikamu chakarurama che β-ME chakawedzerwa, ona danho rekutanga rekuita).
4.The eluent yakawedzerwa zvisizvo.
Zano: Ita shuwa kuti RNase-Yemahara ddH2O inodonhedzwa pakati pechenese column membrane.
5.Iyo chaiyo vhoriyamu yeabsolute ethanol haina kuwedzerwa kuBuffer PSL2 kana Buffer PRW2.
Zvaungaita: Tapota tevera mirairo, wedzera huwandu chaihwo hweabsolute ethanol kuBuffer PSL2 neBuffer PRW2 uye sanganisa zvakanaka kititi isati yashandiswa.
6.Kuwanda kwemuenzaniso wenyama hakuna kufanira.
Zano: Shandisa 50 mg yetishu pa500 μl yeBuffer PSL1.Kushandisa nyama yakawandisa kunoderedza huwandu hweRNA yakabviswa uye kuchena kweiyo RNA inoguma kunodzikiswa zvakare.Isu tinokurudzira zvakasimba kuti yekutanga sampuli dosage haifanire kudarika 50 mg paRNA yekubvisa oparesheni.
7. Vhoriyamu yekuvhota isina kufanira kana kusakwana.
Zano: Vhoriyamu yakasarudzika yekoramu yekuchenesa ndeye 50-200 μl;kana iyo elution mhedzisiro isiri inogutsa, zvinokurudzirwa kuwedzera nguva pakamuri tembiricha mushure mekuwedzera preheated RNase-Mahara ddH.2O, senge 5-10min.
8.Koramu yekuchenesa ine ethanol yakasara mushure mekugeza neBuffer PRW2.
Zano: Kana iyo chubhu isina chinhu iine centrifuged ye1 min uye ichine ethanol yakasara mushure mekugezwa muBuffer PRW2, unogona kuwedzera nguva yechubhu isina chinhu centrifugation kusvika ku2 min, kana kuisa mbiru yekuchenesa pakupisa kwekamuri ye5 min kuti ubvise zvachose ethanol yakasara.
9.Kiti yakashandiswa zvisizvo.
Zano: Kune zvidyarwa zvepolyphenolic polysaccharides, uchishandisa zvakajairika kits sePlant Yese RNA Isolation Kit inogona kusakwanisa kuwana yakakodzera RNA samples.Isu tinokukurudzira kuti ushandise Plant Yese RNA IsolationKit Plus, iyo yakanyatsogadzirirwa polyphenolic polysaccharide chirimwa samples.Kiti yakagadziridzwa kuburitsa RNA kubva ku polyphenol uye polysaccharide chirimwa samples.
OD260/OD280 kukosha kwakaderera
RNA elution ine ddH2O uye inoshandiswa pakuverenga spectrophotometer inoguma yakaderera OD260/OD280 kukosha.Tinokurudzira kushandisa 10 mM Tris-HCl, pH 7.5 (pane RNase-Mahara ddH2O to elute RNA) kuti uwane zvakati tsvikiti zveOD260/OD280, ona “RNA Concentration and Purification Assays” papeji 19.
Iyo RNA yakacheneswa yakaderedzwa
Hunhu hweRNA yakacheneswa hune hukama nezvinhu zvakaita sekuchengetedza sampuli, kusvibiswa kweRNase, uye kunyengera.
Kuongororwa kwezvikonzero zvinowanzoitika:
1.Tissue samples haana kuchengetwa munguva mushure mekuunganidza.
Kurudziro: Kana masampuli ematishu asina kushandiswa munguva mushure mekuunganidzwa, ndapota achengetedze mu nitrogen yemvura pa tembiricha yakaderera pakarepo kana kuti uendese kune -80 ° C kuitira kuchengetedza kwenguva refu mushure mekukurumidza kutonhora mu nitrogen yemvura, kana kunyudza nekukurumidza masampuli muRNA stabilizer RNAlater solution (mhuka samples ).Nekubvisa RNA, edza kushandisa masampula ematishu achangobva kuunganidzwa.
2.Kudzokororwa kwechando uye kunyunguduka kwemasampuli ematishu.
Zvaungaita: Paunenge uchichengeta matishu esampuli, zviri nani kuacheka kuita zvidimbu zvidiki kuti zvichengeteke, uye wotora chikamu chawo paunenge uchiashandisa kudzivirira kuparara kweRNA kunokonzerwa nekuomesa nechando kakawanda uye kunyunguduka kwemasamples acho.
3.RNase inounzwa mukamuri yekuvhiya kana kusapfekwa magirovhosi anoraswa, masiki, nezvimwe.
Zano: RNA kutorwa kwekuyedza kunoitwa zvakanyanya mumabasa akasiyana eRNA, uye tafura yemurabhoritari inofanirwa kucheneswa kusati kwaedzwa, uye magirovhosi nemasks anoraswa anofanira kupfekwa panguva yekuyedza kudzivirira kuparara kweRNA kunokonzerwa nekuunzwa kweRNase zvakanyanya.
4.Iyo reagent yakasvibiswa neRNase panguva yekushandiswa.
Zano: Tsiva nenhevedzano itsva yechirimwa yakazara RNA yekubvisa kits yezviyedzo zvinoenderana.
5.The centrifuge tubes uye pipette mazano anoshandiswa paRNA manipulation akasvibiswa neRNase.
Zvaungaita: Ita shuwa kuti machubhu ecentrifuge, matipi epipette, mapipetti, nezvimwewo zvinoshandiswa muRNA kudhirowa zvese zviri RNase-Mahara.
Mirayiridzo yezvinyorwa:
