Semela Kakaretso ea RNA Isolation Kit Plus Kakaretso ea RNA Purificaiton Kit bakeng sa Semela se ruileng ka Polysaccharides le Polyphenols
Tlhaloso ea Kit:
Katse.No.RE-05021/05022/05024
Bakeng sa tlhoekiso ea kakaretso ea RNA ho tsoa lisampoleng tse akaretsang tsa semela tse nang le likarolo tse phahameng tsa polysaccharide le polyphenol.
Ka potlako ntša RNA ea boleng bo holimo ho lisampole tsa semela tse nang le litaba tse phahameng tsa polysaccharides le polyphenols.
RNase-Free Ho Sebelisa DNA-Cleaning Column
E bonolo - ts'ebetso eohle e phethoa ka mocheso oa kamore
Ka potlako-ts'ebetso e ka phethoa ka metsotso e 30
E bolokehile - ha ho na reagent ea tlhaho e sebelisoang 
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 phahameng kapa polyphenols.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 |
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.
■ E loketse ka ho khetheha bakeng sa tlhoekiso ea RNA ho tloha lisampole tsa limela tsa polysaccharides le polyphenols.
■ 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.
Lihlahisoa tsa lihlahisoa
■ Lisebelisoa tse tlase: motsoako oa pele oa cDNA, RT-PCR, cloning ea molek'hule, Northern Blot, joalo-joalo.
■ Mohlala: Lithishu tsa semela tse ncha kapa tse hatselitsoeng tsa polysaccharides le polyphenols
■ Tekanyetso: 50mg dithishu tsa semela
■ Boholo bo tlamang ba RNA ea kholomo ea tlhoekiso: 80 μg
■ Molumo oa elution: 50-200 μl
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 phahameng 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
Storage le Shelf bophelo
Setsi sena se ka bolokoa likhoeli tse 24 tlas'a maemo a omileng mocheso oa kamore (15-25 ℃);haeba e hloka ho bolokoa nako e telele, e ka bolokoa ka 2–8℃.
Buffer PSL1 e ka behoa ho 4℃ bakeng sa khoeli e le 1 ka mor'a ho eketsa β-mercaptoethanol (ho khothaletsoa ho e kenyelletsa ka nako e tšoanang ea teko).
Kholomo e kentsoe
Ka mor'a hore kholomo e kenngoe, tlhahiso ea RNA e fokotsehile kapa ha e khonehe ho hloekisa RNA, 'me boima ba RNA bo fumanoeng bo tlaase.
Tlhahlobo ea sesosa se tloaelehileng:
1. Likheo tsa lisampole ha lia phethahala.
Ho robeha ha mohlala ha ho etse hore DNA-CLEANING COLUMN e koetsoe ka botlalo, ha e ntse e ama tlhahiso ea RNA le boleng.Re khothaletsa ts'ebetso ea ho sila ka potlako ka naetrojene e lekaneng ea mokelikeli ha o roba lisampole, Leka ho pshatla lerako la seleng ea mohlala, lera la sele le lisele tse ling.Bakeng sa disampole tsa semela tsa polyol polysaccharides, re khothaletsa hore o sebelise Plant Total RNA IOLATION KIT PLUS.
2. Ha o monya sampole e arohaneng ea supernatant e nang le DNA-Cleaning Column, ho ka etsahala hore ebe sele e arohaneng ea precipitate e ka huleloa.
Lisele tse arohaneng tsa lisele tse nkiloeng li tla etsa hore Kholomo ea RNA-FEELA e tla thibeloa ha ts'ebetso ea papatso ea RNA e etsoa (sheba mohato oa 6).Re u khothalletsa hore u be hlokolosi ha u monya supernatant ena ho qoba litšila tsa lisele tse ka monngoang.
3. Chelete ea pele ea mohlala e ngata haholo.
Tšebeliso e feteletseng ea sampole e tla fella ka karohano e sa fellang ea sampole kapa lisele tse sa phethahalang tsa lysis ka Buffer PSL1, e leng se tla fella ka ho thibela kholomo ea tlhoekiso nakong ea tlhoekiso.Plant Total RNA Isolation Kit Mohlala o mong le o mong o hloekisitsoeng o sebetsang ke 50 mg.Bakeng sa disampole tsa semela tsa polyol polysaccharides, re khothaletsa hore o leke Plant Total RNA IOLATION KIT PLUS.
4. Mocheso oa centrifuge o tlase haholo.
Ts'ebetso eohle ea ho itšehla thajana le tlhoekiso ea RNA e etsoa ka mocheso oa kamore (20-25°C), ntle le hore lisele tsa mohlala li robehile ke metsi a naetrojene. Thempereichara ea li-centrifuges tse ling tsa cryogenic e ka tlase ho 20.℃, e ka bakang thibelo ea DNA-Cleaning Column le/kapa RNA-Feela Column.Haeba sena se etsahala, beha mocheso oa centrifuge ho 20-25℃, leetsa bonnete ba hore motsoako oa lysis le/kapa ethanol-e ekelitsoeng supernatant e ne e futhumalitsoe pele 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 etsa bath ea leqhoa le mocheso o tlaase oa centrifugation.
2.RNA e senyehile ka lebaka la ho boloka sampuli e sa lokelang kapa ho boloka nako e telele ea mohlala.
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.Ho se lekane ho arohana ha sampuli le lysis ho lebisa ho thibela 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 ho hloekisa.
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 o sa nepahalang oa ho hlakisa 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 ka mocheso oa kamore ka mor'a ho eketsa RNase-Free ddH2O ea preheated, joalo ka 5-10min.
8.Kolumo ea tlhoekiso e na le masala a ethanol ka mor'a ho hlatsoa le BufferPRW2.
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 'me e sebelisetsoa ho bala spectrophotometer e fella ka boleng bo tlaase ba OD260/OD280.Re khothaletsa ho sebelisa 10 mM Tris-HCl, pH 7.5 (ho e-na le RNase-Free ddH2O ho elute RNA) ho fumana boleng bo batlang bo nepahetse ba 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:
Plant Total RNA Isolation Kit Plus Instruction Manual
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