Rnase Inhibitor (Glycerol mahala)
Murine RNase inhibitor ke recombinant murine RNase inhibitor e hlahisitsoeng le ho hloekisoa ho tsoa ho E.coli.E tlama ho RNase A, B kapa C ka karolelano ea 1: 1 ka ho kopanya ho se nang covalent, kahoo e thibela mosebetsi oa li-enzyme tse tharo le ho sireletsa RNA ho senyeha.Leha ho le joalo, ha e sebetse khahlanong le RNase 1, RNase T1, S1 Nuclease, RNase H kapa RNase ho tloha Aspergillus.Murine RNase inhibitor e ile ea lekoa ke RT-PCR, RT-qPCR le IVT mRNA, 'me e ne e lumellana le mefuta e fapaneng ea khoebo ea Reverse transcriptases, DNA polymerases le RNA polymerase.
Ha ho bapisoa le li-inhibitors tsa RNase tsa batho, murine RNase inhibitor ha e na li-cysteine tse nang le maikutlo a matla haholo ho oxidation e bakang ho se sebetse ha inhibitor.Seo se etsa hore e tsitsitse maemong a tlase a DTT (tlase ho 1 mM).Tšobotsi ena e etsa hore e tšoanelehe ho sebelisoa karabelong moo likhakanyo tse phahameng tsa DTT li leng khahlanong le karabelo (mohlala, nako ea 'nete RT-PCR).
Akopo
Sehlahisoa sena se ka sebelisoa haholo tekong efe kapa efe moo tšitiso ea RNase e ka khonehang ho qoba ho senyeha ha RNA, joalo ka:
1.Tlhaloso ea pele ea cDNA, RT-PCR, RT-qPCR, joalo-joalo;
2.Sireletsa RNA ho tloha ho senyeha ha in-vitro transcription/phetolelo (mohlala, viral replication in vitro);
3.Ho thibela mosebetsi oa RNase nakong ea ho itšehla thajana le ho hloekisoa ha RNA.
Maemo a polokelo
Boloka ho -25～-15℃;
Lipotoloho tsa ho qhibiliha ka leqhoa ≤ linako tse 5;
E sebetsa lilemo tse 1.
Tlhaloso ea yuniti
Yuniti e le 'ngoe e hlalosoa e le palo ea RNase Inhibitor e hlokahalang ho thibela tšebetso ea 5ng ea RNase A ka 50%.
Boima ba molek'hule
RNase Inhibitor (e se nang Glycerol) ke protheine ea 50 kDa.
Taolo ea boleng
Exonuclease Ketsahalo:
Ho kenngoa ha 40 U ea enzyme e nang le 1 μg λ-Hind III cheka DNA bakeng sa lihora tse 16 ho 37 ° C ho entse hore ho se be le ho senyeha ho bonahalang ha DNA joalokaha ho laetsoe ke gel electrophoresis.
Ts'ebetso ea Endonuclease:
Ho kenngoa ha 40 U ea enzyme e nang le 1 μg λ DNA bakeng sa lihora tse 16 ho 37 ° C ho entse hore ho se be le ho senyeha ho bonahalang ha DNA joalokaha ho laetsoe ke gel electrophoresis.
Ho thala Ketsahalo:
Ho kenngoa ha 40 U ea enzyme e nang le 1 μg pBR322 bakeng sa lihora tse 16 ho 37 ℃ ho entse hore ho se be le ho senyeha ho bonahalang ha DNA joalokaha ho laetsoe ke gel electrophoresis.
RNase Ketsahalo:
Ho kenngoa ha 40 U ea enzyme e nang le 1.6 μg MS2 RNA bakeng sa lihora tse 4 ho 37 ℃ ho entse hore ho se be le ho senyeha ho bonahalang ha RNA joalokaha ho laetsoe ke gel electrophoresis.
E.coli DNA:
40 U ea Enzyme e bonoa ke TaqMan qPCR.DNA ea E.coli ke ≤ 0. 1pg/40U.
Notes
1.U se ke ua sisinyeha kapa ua sisinya ka matla ho thibela ho sebetsa ha enzyme.
2.RNase inhibitor e sebetsa ka mocheso ho tloha 25 ℃ ho ea ho 55 ℃ 'me ha e sebetse ho ≥65℃.
3.Mosebetsi oa RNase H, RNase 1, le RNase T1 ha oa thibeloa.
4. Thepa ea pH bakeng sa ho thibela ts'ebetso ea RNase e pharaletse (e sebetsa ho pH 5-9), e bonts'a ts'ebetso e phahameng ho pH 7-8.
