I-Rnase Inhibitor

Inombolo yekati:HC2010A

Iphakheji: 2KU/10KU/100KU

I-Murine RNase inhibitor iyi-recombinant murine RNase inhibitor evezwe futhi yahlanzwa ku-E.coli.

Thumela Uphenyo

Incazelo Yomkhiqizo

Imininingwane yomkhiqizo

I-Murine RNase inhibitor iyi-recombinant murine RNase inhibitor evezwe futhi yahlanzwa ku-E.coli.Ibophezela ku-RNase A, B noma C ngesilinganiso esingu-1:1 ngokusebenzisa i-non-covalent bonding, ngaleyo ndlela ivimbele umsebenzi wama-enzyme amathathu futhi ivikele i-RNA ekonakalisweni.Nokho, Ayisebenzi uma iqhathaniswa ne-RNase 1, RNase T1, S1 Nuclease, RNase H noma i-RNase evela ku-Aspergillus.I-Murine RNase inhibitor yahlolwa yi-RT-PCR, RT-qPCR kanye ne-IVT mRNA, futhi yayihambisana nezinhlobonhlobo ze-Reverse transcriptases zezentengiselwano, ama-DNA polymerases nama-polymerase e-RNA.

Uma kuqhathaniswa nama-RNase inhibitors abantu, i-murine RNase inhibitor ayinawo ama-cysteines amabili azwela kakhulu ku-oxidation ebangela ukungasebenzi kwe-inhibitor.Lokho kuyenza izinze ekugxilweni okuphansi kwe-DTT (ngaphansi kuka-1 mM).Lesi sici siyenza ifanelekele ukusetshenziswa ekuphenduleni lapho ukugxila okuphezulu kwe-DTT kuphambene nokusabela (isb. I-Real-time RT-PCR).

Okwedlule: I-Hotstart Taq DNA Polymerase HC1012A

Olandelayo: I-Hs Taq DNA Polymerase (i-Glycerol-free)

Aisicelo

Lo mkhiqizo ungasetshenziswa kakhulu kunoma yikuphi ukuhlola lapho ukuphazamiseka kwe-RNase kungenzeka khona ukuze kugwenywe ukucekelwa phansi kwe-RNA, okufana nalokhu:

1.Ukuhlanganiswa komucu wokuqala we-cDNA, i-RT-PCR, i-RT-qPCR, njll.

2.Ivikela i-RNA ekonakalisweni ngesikhathi sokuloba/ukuhumusha kwe-in vitro (isb., uhlelo lokuphindaphinda kwamagciwane ku-vitro).

3.Ukuvinjelwa komsebenzi we-RNase ngesikhathi sokuhlukaniswa kwe-RNA nokuhlanzwa.

Izimo Zokugcina

Umkhiqizo ungagcinwa ku -25～- 15 ℃, usebenze iminyaka emi-2.

Ibhafa yesitoreji

50 mM KCl, 20 mM HEPES-KOH (pH 7.6, 25 ℃), 8 mM DTT kanye no-50% we-glycerol.

Incazelo yeyunithi

Inani le-murine RNase inhibitor elidingekayo ukuze kuvinjwe umsebenzi we-5ng we-ribonuclease A ngo-50% lichazwe njengeyunithi eyodwa (U).

Isisindo samangqamuzana amaprotheni

Isisindo samangqamuzana e-murine RNase inhibitor singama-50 kDa.

Ikhwalithi yokulawula

I-Exonuclease Umsebenzi:

I-40 U ye-murine RNase inhibitor ene-1 μg λ -Hind III yokugaya i-DNA ku-37℃ amahora angu-16 ayikhiqizi ukonakala njengoba kunqunywe i-agarose gel electrophoresis.
Umsebenzi we-Endonuclease:

I-40 U ye-murine RNase inhibitor ene-1μ g λ DNA ku-37℃ amahora angu-16 ayikhiqizi njengoba kunqunywe i-agarose gel electrophoresis.

Ukudlala Umsebenzi:

I-40U ye-murine RNase inhibitor ene-1μ g pBR322 ku-37℃ amahora angu-16 ayikhiqizi ukonakala njengoba kunqunywa i-agarose gel electrophoresis.

RNase Umsebenzi:

I-40U ye-murine RNase inhibitor ene-1.6μ g MS2 RNA ye-4h ku-37℃ ayikhiqizi ukonakala njengoba kunqunywe i-agarose gel electrophoresis.

E.coli DNA:

I-40 U ye-murine RNase inhibitor ihlolelwa ukuba khona kwe-E. coli genomic DNA kusetshenziswa i-TaqMan qPCR eneziqalo eziqondile ze-E. coli 16S rRNA locus.Ukungcola kwe-E. coli genomic DNA kungu-≤ 0.1 pg/40 U.

Nothis

I-1.I-oscillation enobudlova noma ukugqugquzela kuzoholela ekusebenzeni kwe-enzyme.

2.Izinga lokushisa elilungile lale inhibitor lalingu-25-55 ℃, futhi Lalingasebenzi ku-65℃ nangaphezulu.

3.Imisebenzi ye-RNase H, i-RNase 1 ne-RNase T1 ayizange ivinjwe i-murine RNase inhibitor.

4.Ukuvinjelwa komsebenzi we-RNase kutholwe kuhlu olubanzi lwe-pH (i-pH 5-9 yonke yayisebenza), futhi umsebenzi ophakeme kakhulu wabonwa ku-pH 7-8.

5.Njengoba ama-ribonuclease ngokuvamile agcina umsebenzi ngaphansi kwezimo ze-denaturing, kufanele kuqashelwe ukugwema ukukhipha ama-molecule e-RNase Inhibitor ahlanganiswe ne-ribonuclease.Ukuze uvimbele ukukhishwa kwe-ribonuclease esebenzayo, izinga lokushisa elingaphezu kuka-50 °C kanye nokugxila okuphezulu kwe-urea noma amanye ama-denaturing agents kufanele kugwenywe.