Mali a RNA Isolation Kit
Tlhaloso ea Kit:
Katse.No.RE-04011/04013
Bakeng sa tlhoekiso e felletseng ea RNA maling a felletseng 104 ≤ Lisele tse tšoeu tsa mali ≤ 107
Ka potlako arola le ho hloekisa mali a RNA liseleng tse tšoeu tsa mali.
-Ha ho hlokahale hore u tšoenyehe ka ho senyeha ha RNA.Thepa eohle ha e na RNase-Free
-E bonolo - ts'ebetso eohle e phethoa ka mocheso oa kamore
-Ka potlako-ts'ebetso e ka phethoa ka metsotso e 20
-Sehlahisoa se phahameng sa RNA: Kholomo ea RNA feela le foromo e ikhethang e ka hloekisa RNA hantle
-Sireletsehile-ha ho na reagent ea tlhaho e sebelisoang
-Mehlala e meholo ea ho sebetsa - ho fihlela ho 200μl lisampole li ka sebetsoa nako le nako.
-Boleng bo phahameng-RNA e hloekisitsoeng e hloekile haholo, ha e na liprotheine le litšila tse ling, 'me e ka kopana le mekhoa e fapaneng ea liteko tse tlaase.
Litlhaloso
Thepa e nka "spin column le formula" e ntlafalitsoeng ke k'hamphani ea rona, e ka ntšang RNA ea boleng bo holimo le ea boleng bo holimo maling a anticoagulated.Setsi sena se fana ka lisele tse khubelu tsa mali lysate (Buffer RCL), e ka khonang ho ntša lisele tse khubelu tsa mali kapele le ka katleho le ho boloka lisele tse tšoeu tsa mali.DNA-Cleaning Colunm e sebetsang hantle e ka arola habonolo supernatant le cell lysates le adsorb le ho tlosa genomic DNA.Ts'ebetso e bonolo ebile e boloka nako;Kholomo ea RNA feela e ka tlama RNA ka nepo, 'me ka foromo e ikhethang, e ka sebetsana le lisampole tse ngata ka nako e le ngoe.
Tsamaiso eohle ea RNase-Free e etsa hore RNA e nkiloeng e se ke ea senyeha;Buffer RW1 le Buffer RW2 buffer system e etsa hore RNA e fumanoeng e se na protheine, DNA, ion le tšilafalo ea motsoako oa organic.
Litaba tsa Kit
| Mali Kakaretso ea RNA Isolation Kit | ||
| Sebopeho sa Kit | RE-04011 | RE-04013 |
| 50 linako tse ling | 200 linako tse ling | |
| Buffer RCL (10×) | 52.5 mL | 210mL |
| Buffer BRL1* | 30 ml | 120mL |
| Buffer BRL2 | 18mL | 66ml |
| Buffer RW1* | 25mL | 100mL |
| Buffer RW2 | 24mL | 96 ml |
| RNase-Free ddH2 O | 10mL | 40mL |
| Lekholomo la RNA feela | 50 lihlopha | 200 lihlopha |
| DNA-Ho hloekisa Column | 50 lihlopha | 200 lihlopha |
| tataiso | 1 khopi | 1 khopi |
Likarolo le melemo
-Ha ho hlokahale hore u tšoenyehe ka ho senyeha ha RNA.Thepa eohle ha e na RNase-Free.
-E bonolo - ts'ebetso eohle e phethoa ka mocheso oa kamore.
-Ka potlako-ts'ebetso e ka phethoa ka metsotso e 20.
-Sehlahisoa se phahameng sa RNA: Kholomo ea RNA feela le foromo e ikhethang e ka hloekisa RNA hantle.
-Sireletsehile-ha ho na reagent ea tlhaho e sebelisoang.
-Mehlala e meholo ea ho sebetsa - ho fihlela ho 200μl lisampole li ka sebetsoa nako le nako.
-Boleng bo phahameng-RNA e hloekisitsoeng e hloekile haholo, ha e na liprotheine le litšila tse ling, 'me e ka kopana le mekhoa e fapaneng ea liteko tse tlaase.
Litekanyetso tsa kit
Sesebelisoa sa Kit:
E loketse ho ntšoa le ho hloekisoa ha RNA kaofela ho tsoa maling a mammalian kaofela.
Phallo ea mosebetsi
Maemo a polokelo
Buffer RCL (10 ×) e lokela ho bolokoa ho 2-8 ℃;likarolo tse ling tsa kit li ka bolokoa mocheso oa kamore (15-25 ℃) tlas'a maemo a omileng, 'me li ka bolokoa likhoeli tse 12.Buffer BRL1 e ka bolokoa ho 4 ℃ bakeng sa khoeli e le 'ngoe ka mor'a ho eketsa β-mercaptoethanol (ho ikhethela) .
Tlhokomeliso: Haeba e bolokiloe mochesong o tlase, tharollo e atisa ho ba le pula.Etsa bonnete ba hore o beha tharollo ka har'a kit mocheso oa kamore nako e itseng pele o sebelisoa.Haeba ho hlokahala, e chese pele ka bateng ea metsi ea 37 ° C metsotso e 10 ho qhala metsi, 'me u kopanye hantle pele u sebelisoa.
Litaelo tsa tlhahlobo ea mathata
The following is an analysis of the problems that might be encountered in the extraction of viral RNA. We wish it would be helpful to your experiment. In addition, for other experimental or technical problems other than operating instructions and problem analysis, we have dedicated technical support to help you. Contact us if you need at : 028-83361257or E-mail：Tech@foregene.com。
Ha ho RNA e ka ntšoang kapa chai ea nucleic acid e tlase
Hangata ho na le lintlha tse ngata tse amang katleho ea ho hlaphoheloa, joalo ka: sampole ea litaba tsa RNA, mokhoa oa ts'ebetso, bophahamo ba molumo, joalo-joalo.
Tlhahlobo ea lisosa tse tloaelehileng:
1.Bate ea leqhoa kapa mocheso o tlase (4 ° C) centrifugation nakong ea ts'ebetso.
Tlhahiso: Mocheso oa kamore (15-25 ° C) o sebetsa, ha ho mohla ho hlatsoa leqhoa le mocheso o tlase oa centrifuge.
2. Polokelo e sa nepahalang ea mohlala kapa polokelo ea mohlala nako e telele haholo.
Tlhahiso: Boloka lisampole ho -80 ° C kapa u hatsele ka har'a metsi a naetrojene, 'me u qobe tšebeliso ea ho qhibiliha khafetsa;leka ho sebelisa lisampole tse sa tsoa bokelloa bakeng sa ho ntša RNA.
3.Sample e sa lekaneng ea lysis
Keletso: Ka kopo etsa bonnete ba hore sampole le tharollo e sebetsang (Linear Acrylamide) li tsoakoe ka botlalo 'me li kentsoe metsotso e 10 mocheso oa kamore (15-25 ° C)
4.The eluent e kentsoe ka phoso
Keletso: Etsa bonnete ba hore RNase-Free ddH2O e kentsoe bohareng ba lera la kholomo ea tlhoekiso.
5.Molumo o sa nepahalang oa ethanol ea anhydrous ho Buffer viRW2
Tlhahiso: Ka kopo latela litaelo, eketsa molumo o nepahetseng oa ethanol ea anhydrous ho Buffer viRW2 'me u li kopanye hantle pele u sebelisa kit.
6.Tsebeliso e fosahetseng ea sampole.
Tlhahiso: 200µl ea sampole ka 500μl ea Buffer viRL.Bophahamo bo feteletseng ba sampole bo tla fella ka ho fokotseha ha sekhahla sa ho ntšoa ha RNA.
7.Molumo o sa nepahalang oa tlhahiso kapa tlhahiso e sa fellang.
Tlhahiso: Molumo o hlakileng oa kholomo ea tlhoekiso ke 30-50μl;haeba phello ea elution e sa khotsofatse, ho kgothaletswa ho eketsa RNase-Free ddH e futhumetseng2O le ho eketsa nako ea ho beha mocheso oa kamore, joalo ka 5-10min
8.Kolumo ea tlhoekiso e na le masalla a ethanol ka mor'a ho hlatsoa Buffer viRW2.
Tlhahiso: Haeba ethanol e ntse e sala ka mor'a hore e hlatsoe ka Buffer viRW2 le centrifugation e se nang letho bakeng sa 2min, kholomo ea tlhoekiso e ka siuoa mocheso oa kamore bakeng sa 5min ka mor'a hore ho be le letho-tube centrifugation ho tlosa ethanol e setseng ka botlalo.
Ho senyeha ha limolek'hule tsa RNA tse hloekisitsoeng
Boleng ba RNA e hloekisitsoeng bo amana le lintlha tse kang polokelo ea sampuli, tšilafalo ea RNase, le ts'ebetso.
Tlhahlobo ea lisosa tse tloaelehileng:
1.Lisampole tse bokeletsoeng ha lia ka tsa bolokoa ka nako.
Tlhahiso: Haeba sampole e sa sebelisoe ka nako kamora ho bokelloa, ka kopo e boloke ho -80 ℃ kapa metsi a naetrojene hang-hang.Bakeng sa ho ntša limolek'hule tsa RNA, leka ho sebelisa lisampole tse sa tsoa bokelloa neng kapa neng ha ho khoneha.
2.Collected disampole ba ne ba hoama le ho qhibiliha khafetsa.
Tlhahiso: Qoba ho qhoqhoa khafetsa le ho qhibiliha (e seng ho feta hanngoe) nakong ea ho bokella le ho boloka sampole, ho seng joalo chai ea nucleic acid e tla fokotseha.
3.RNase e ile ea hlahisoa ka kamoreng ea ho buuoa kapa ho ne ho se na liatlana tse lahliloeng, limaske, joalo-joalo.
Tlhahiso: Ho ntšoa ha liteko tsa limolek'hule tsa RNA ho etsoa hantle ka phaposing e arohaneng ea tšebetso ea RNA, 'me tafole ea liteko e hloekisoa pele ho teko.Apara liatlana le limaske tse lahloang nakong ea liteko ho qoba ho senyeha ha RNA ho bakoang ke kenyelletso ea RNase.
4.The reagent e silafalitsoe ke RNase nakong ea tšebeliso.
Tlhahiso: Kenya sebaka ka Viral RNA Isolation Kit e ncha bakeng sa liteko tse amanang.
5. Tšilafalo ea RNase ea liphaephe tsa centrifuge, malebela a pipette, joalo-joalo. Tlhahiso: Etsa bonnete ba hore li-tubes tsa centrifuge, malebela a pipette, le lipeipi kaofela ha li na RNase.
Limolek'hule tsa RNA tse hloekisitsoeng li ile tsa ama liteko tse tlase
Limolek'hule tsa RNA tse hloekisitsoeng ke kholomo ea tlhoekiso li tla ama liteko tse tlase tsa metsi haeba ho na le li-ion tsa letsoai kapa liprotheine tse ngata haholo, joalo ka: reverse transcription, Northern Blot, joalo-joalo.
1.Ho na le li-ion tse setseng tsa letsoai ka har'a limolek'hule tsa eluted RNA.
Keletso: Etsa bonnete ba hore molumo o nepahetseng oa ethanol ea anhydrous e kentsoe Buffer viRW2, 'me u hlatsoe kholomo ea tlhoekiso habeli ho latela lebelo le nepahetseng la centrifugation litaelong tsa ts'ebetso; Haeba ho ntse ho e-na le li-ion tsa letsoai tse setseng, u ka eketsa Buffer viRW2 kholomong ea tlhoekiso, 'me u e tlohele mocheso oa kamore bakeng sa 5min.Ebe u etsa centrifugation ho tlosa silafatso ea ion ea letsoai ka tekanyo e kholo ka ho fetisisa
2.Ho na le ethanol e setseng ka har'a limolek'hule tsa RNA
Tlhahiso: hang ha o netefatsa hore litšiea tsa tlhoekiso li hlatsoitsoe ke Buffer viRW2, etsa centrifugation e se nang letho ho latela lebelo la centrifugal ho litaelo tsa ts'ebetso.Haeba ho ntse ho e-na le ethanol e setseng, e ka tloheloa metsotso e 5 mocheso oa kamore ka mor'a hore ho be le centrifugation e se nang letho ho tlosa ethanol e setseng ka tekanyo e kholo ka ho fetisisa.
Litaelo:
Viral RNA Isolation Kit Instruction Manual
