Semela Kakaretso ea RNA Isolation Kit Kakaretso ea RNA Purificaiton Kit bakeng sa Semela se Tlase ho Polysaccharides le Polyphenols
Litlhaloso
50 Preps, 200 Preps
Thepa e sebelisa "spin column le formula" e ntlafalitsoeng ke Foregene, e ka ntšang RNA ea boleng bo holimo le ea boleng bo holimo ho lithane tse fapaneng tsa semela tse nang le li-polysaccharides tse tlase le polyphenols.Bakeng sa lisampole tsa semela tse nang le li-polysaccharides tse phahameng kapa polyphenols, ho khothaletsoa ho sebelisa Plant Total RNA Isolation Plus Kit ho fumana liphetho tse ntle tsa ho ntšoa ha RNA.Kit e fana ka kholomo ea DNA-Cleaning e ka tlosang DNA ea genomic habonolo ho supernatant le lysate ea lisele.Kholomo ea RNA feela e ka tlama RNA ka nepo.Kit e ka sebetsana le palo e kholo ea lisampole ka nako e le 'ngoe.
Tsamaiso eohle ha e na RNase, kahoo RNA e hloekisitsoeng e ke ke ea senyeha.Buffer PRW1 le Buffer PRW2 e ka etsa bonnete ba hore RNA e fumanoeng ha e silafatsoe ke liprotheine, DNA, ions, le metsoako ea tlhaho.
Likarolo tsa Kit
| Buffer PSL1, Buffer PS, Buffer PSL2 |
| Buffer PRW1, Buffer PRW2 |
| RNase-Free ddH2O, DNA-Cleaning Column |
| RNA-Feela Kholomo |
| Litaelo |
Likarolo le melemo
■ Ts'ebetso ka mocheso oa kamore (15-25 ℃) ho pholletsa le ts'ebetso eohle, ntle le ho hlatsoa leqhoa le mocheso o tlaase oa centrifugation.
■ Complete Kit RNase-Free, ha ho hlokahale hore u tšoenyehe ka ho senyeha ha RNA.
■ DNA-Cleaning Column ka ho khetheha e tlama ho DNA, e le hore kit e ka tlosa tšilafalo ea DNA ea genomic ntle le ho eketsa DNase.
■ Poelo e phahameng ea RNA: Kholomo ea RNA feela le foromo e ikhethang e ka hloekisa RNA hantle.
■ Lebelo le potlakileng: ho bonolo ho sebetsa mme le ka phethoa nakong ea metsotso e 30.
■ Tšireletseho: ha ho na reagent ea tlhaho e hlokahalang.
■ Boleng bo phahameng: Likaroloana tsa RNA tse hloekisitsoeng li hloekile haholo, ha li na protheine le litšila tse ling, 'me li ka kopana le mekhoa e fapaneng ea liteko tse tlase.
Sesebelisoa sa Kit
E loketse ho ntšoa le ho hloekisoa ha kakaretso ea RNA ho tloha lisampoleng tse foreshe kapa tse hatselitsoeng (haholo-holo mahlaku a mahlaku a semela) a nang le polysaccharide e tlase le polyphenol.
Phallo ea mosebetsi
Setšoantšo
Plant Total RNA Isolation Kit Plus e sebelitse 50mg ea makhasi a macha a polysaccharides le polyphenols, 'me 5% e hloekisitsoeng ea RNA e ile ea lekoa ke electrophoresis.
1: Banana
2: Ginkgo
3: K'hothone
4: Kharenate
Polokelo le shelf bophelo
Thepa e ka bolokoa likhoeli tse 12 mocheso oa kamore (15-25 ℃) sebakeng se omileng, le 2-8 ℃ nako e telele (likhoeli tse 24).
Buffer PSL1 e ka bolokoa ho 4 ℃ bakeng sa khoeli e le 'ngoe ka mor'a ho eketsa 2-hydroxy-1-ethanethiol(khetho).
Tataiso ea Tlhahlobo ea Mathata
Tlhahlobo e latelang ea mathata ao u ka kopanang le ona ho onaKakaretso ea SemelaRNA 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.
The spin column e koalehile
Ho thibela mokokotlo oa mokokotlo ho tla etsa hore lihlahisoa tsa RNA li fokotsehe kapa li se ke tsa hloekisoa ho fumana RNA, 'me boleng ba RNA e fumanoeng bo tla ba tlaase.
Tlhahlobo ea Sesosa se Tloaelehileng:
1. Mohlala ha o robehe ka ho feletseng.
Karohano e sa phethahalang ea sampole e ka thibela DNA-Cleaning Column, e ka amang hape tlhahiso ea RNA le boleng.Re khothaletsa hore ha u etsa karohano ea sampole, u potlakele ho sila ka bongata bo lekaneng ba naetrojene ea metsi ho pshatla lithane tse kang mabota a lisele le lisele tsa lisampole ka hohle kamoo ho ka khonehang.Bakeng sa disampole tsa semela tsa polyphenol polysaccharides, re khothaletsa hore o sebelise Plant Total RNA Isolation Kit Plus.
2.Aspirate the DNA-Cleaning Column itšehla supernatant, aspirate ka khoneha sele maloanlahla pellet.
Aspirated cell debris pellet e ka koala Kholomo ea RNA-feela nakong ea mekhoa ea papatso ea RNA (sheba mohato oa 5 oa ts'ebetso, mohato oa 6 oa mokhoa oa polysaccharide polyphenol).Re khothaletsa hore motho a be hlokolosi ha a lakatsa ho qoba matlakala a liseleng.
3. Palo ea pele ea sampole e kholo haholo.
Tšebeliso e feteletseng ea sampole e tla fella ka karohano e sa fellang ea sampole kapa lysis e sa fellang ea lisele ka Buffer PRL1 kapa Buffer PSL1, e leng se tla fella ka kholomo e koetsoeng ea tlhoekiso bakeng sa ts'ebetso ea tlhoekiso.Plant Total RNA Isolation Kit e na le boholo ba pele ba 50 mg ka tlhoekiso e le 'ngoe ea sampole e sebelitsoeng.Bakeng sa disampole tsa semela tsa polyphenol polysaccharides, re khothaletsa hore o leke Plant Total RNA Isolation Kit Plus.
4. Mocheso oa centrifuge o tlase haholo.
Ho itšehla thajana le tlhoekiso ea RNA e felletseng ntle le tšitiso ea naetrojene ea metsi ea sampole, mehato eohle e etsoa ka mocheso oa kamore (20-25 ° C).Li-centrifuge tse ling tsa mocheso o tlase li na le mocheso o ka tlase ho 20 °C, e leng se ka bakang tšitiso ho Kholomo ea DNA-Cleaning le/kapa Kholomo ea RNA-feela.Haeba sena se etsahala, beha mocheso oa centrifuge ho 20-25 ° C 'me u futhumale pele motsoako oa lysis le / kapa ho eketsoa ha karohano ea ethanol supernatant ho 37 °C.
Ha ho RNA e nkiloeng kapa tlhahiso ea RNA e tlase
Hangata ho na le lintlha tse ngata tse amang ts'ebetso ea ho hlaphoheloa, joalo ka: sampole ea litaba tsa RNA, mokhoa oa ts'ebetso, bophahamo ba molumo, jj.
Tlhahlobo ea lisosa tse tloaelehileng tse latelang:
1.Ho hlatsoa leqhoa kapa mocheso o tlaase (4 ° C) centrifugation e ne e etsoa nakong ea ts'ebetso.
Tlhahiso: Sebetsa ka mocheso oa kamore (15-25 ° C) ts'ebetsong eohle, u se ke ua hlatsoa leqhoa le mocheso o tlaase oa centrifugation.
2.RNA e senyehile ka lebaka la paballo e fosahetseng ea sampole kapa poloko ea nako e telele ea sampole.
Keletso: lisampole tse sa tsoa bokelloa li lokela ho hoamisoa ka potlako ka har'a metsi a naetrojene, ebe li bolokoa ho -80°C nako e telele, ho qoba ho hatsetsoa khafetsa le ho qhibiliha ha lisampole;kapa hang-hang kenya lisampole ka har'a tharollo ea RNA stabilizer RNAlater (lisampole tsa liphoofolo).
3.Karohano e sa lekaneng ea sampole le lysis e lebisa ho thibeloa ha kholomo ea tlhoekiso.
Tlhahiso: Ha u sila lesela, ka kopo etsa bonnete ba hore lesela le fatše ka ho lekaneng, 'me ka potlako le fetisetse ho Buffer PSL1 e lokiselitsoeng esale pele (tiisa hore karolo e nepahetseng ea β-ME e kentsoe, sheba mohato oa 1 oa mokhoa).
4.The eluent e kentsoe ka phoso.
Tlhahiso: Etsa bonnete ba hore RNase-Free ddH2O e rothela bohareng ba lera la tlhoekiso.
5.Bolumo e nepahetseng ea ethanol e felletseng ha ea eketsoa ho Buffer PSL2 kapa Buffer PRW2.
Tlhahiso: Ka kopo latela litaelo, eketsa molumo o nepahetseng oa absolute ethanol ho Buffer PSL2 le Buffer PRW2 'me u kopanye hantle pele khiti e sebelisoa.
6.Bongata ba sampole ea linama ha boa lokela.
Tlhahiso: Sebelisa 50 mg ea lisele ho 500 μl ea Buffer PSL1.Ho sebelisa lisele tse ngata haholo ho tla fokotsa palo ea RNA e nkiloeng, 'me bohloeki ba RNA e hlahisoang le bona bo tla fokotseha.Re khothaletsa ka matla hore tekanyetso ea pele ea sampole e se ke ea feta 50 mg ka ts'ebetso ea ho ntša RNA.
7. Molumo oa lentsoe o sa nepahalang kapa tlhahiso e sa fellang.
Tlhahiso: Molumo o hlakileng oa kholomo ea tlhoekiso ke 50-200 μl;haeba phello ea elution e sa khotsofatse, ho kgothaletswa ho eketsa nako mocheso oa kamore ka mor'a ho eketsa RNase-Free ddH2O, joalo ka 5-10min.
8.Kolumo ea tlhoekiso e na le masalla a ethanol ka mor'a ho hlatsoa ka Buffer PRW2.
Tlhahiso: Haeba tube e se nang letho e le centrifuged bakeng sa 1 min 'me ho ntse ho e-na le ethanol e setseng ka mor'a ho hlatsoa Buffer PRW2, u ka eketsa nako ea centrifugation ea tube e se nang letho ho 2 min, kapa u behe mohala oa tlhoekiso mocheso oa kamore bakeng sa 5 min ho tlosa ethanol e setseng ka botlalo.
9. Thepa e sebelisitsoe hampe.
Tlhahiso: Bakeng sa lisampole tsa limela tsa polyphenolic polysaccharides, ho sebelisa lisebelisoa tse tloaelehileng tse kang Plant Total RNA Isolation Kit ho ka 'na ha se khone ho fumana lisampole tse loketseng tsa RNA.Re u khothaletsa ho sebelisa Plant Total RNA IsolationKit Plus, e etselitsoeng lisampole tsa semela sa polyphenolic polysaccharide ka ho khetheha.Kit e etselitsoeng ho hula RNA ho tsoa ho disampole tsa semela sa polyphenol le polysaccharide.
OD260/OD280 boleng bo tlase
Tlhahiso ea RNA e nang le ddH2O le ho sebelisoa bakeng sa ho bala spectrophotometer ho fella ka boleng bo tlase ba OD260/OD280.Re khothaletsa ho sebelisa 10 mM Tris-HCl, pH 7.5 (ho ena le RNase-Free ddH2O to elute RNA) ho fumana litekanyetso tse nepahetseng tsa OD260/OD280, bona “RNA Concentration and Purification Assays” leqepheng la 19.
RNA e hloekisitsoeng e senyehile
Boleng ba RNA e hloekisitsoeng bo amana le lintlha tse kang ho boloka sampole, tšilafalo ea RNase, le ho qhekella.
Tlhahlobo ea lisosa tse tloaelehileng:
Li-sampuli tsa 1.Tissue ha lia ka tsa bolokoa ka nako ka mor'a ho bokella.
Keletso: Haeba lisampole tsa linama li sa sebelisoe ka nako ka mor'a ho bokelloa, ka kopo li boloke ka har'a naetrojene e nang le metsi ka mocheso o tlase hang-hang kapa u li fetisetse ho -80 ° C bakeng sa polokelo ea nako e telele ka mor'a ho hatseha ka potlako ka naetrojene e nang le metsi, kapa hang-hang u qoelise lisampole ka RNA stabilizer RNAlater solution (li-sampuli tsa liphoofolo ).Bakeng sa ho ntša RNA, leka ho sebelisa lisampole tsa linama tse sa tsoa bokelloa.
2.Mahatsetso a phetoang le ho qhibiliha ha lisampole tsa linama.
Tlhahiso: Ha u boloka lisampole tsa linama, ho molemo hore u li khaole likotoana tse nyenyane bakeng sa ho bolokoa, ’me u ntše karolo e ’ngoe ea tsona ha u li sebelisa ho qoba ho senyeha ha RNA ho bakoang ke ho ba leqhoa khafetsa le ho qhibiliha ha lisampole.
3.RNase e hlahisoa ka phapusing ea ts'ebetso kapa e sa roaloe liatlana tse lahliloeng, limaske, jj.
Tlhahiso: Liteko tsa ho ntša RNA li etsoa hantle ka ho fetisisa mesebetsing e arohaneng ea RNA, 'me tafole ea laboratori e lokela ho hloekisoa pele ho teko,' me li-gloves le limaske tse lahloang li lokela ho apesoa nakong ea teko ho qoba ho senyeha ha RNA ho bakoang ke ho kenngoa ha RNase ka tekanyo e kholo ka ho fetisisa.
4.The reagent e silafalitsoe ke RNase nakong ea tšebeliso.
Tlhahiso: Fetosa ka letoto le lecha la lisebelisoa tsa RNA tsa limela bakeng sa liteko tse amanang.
5. Li-tubes tsa centrifuge le malebela a pipette a sebelisoang bakeng sa ho fetola RNA li silafalitsoe ke RNase.
Tlhahiso: Etsa bonnete ba hore li-tubes tsa centrifuge, malebela a pipette, pipette, joalo-joalo tse sebelisoang ho ntša RNA kaofela ha li na RNase.
Litaelo:
