I-Rnase Inhibitor

Inombolo yekati:HC2010A

Iphakheji: 2KU/10KU/100KU

I-Murine RNase inhibitor yi-recombinant murine RNase inhibitor echazwe kwaye ihlanjululwe kwi-E.coli.

Thumela uMbuzo

Ingcaciso yeMveliso

Iinkcukacha zemveliso

I-Murine RNase inhibitor yi-recombinant murine RNase inhibitor echazwe kwaye ihlanjululwe kwi-E.coli.Ibophelela kwi-RNase A, B okanye C kwi-1: umlinganiselo we-1 ngokusebenzisa i-non-covalent bonding, ngaloo ndlela inqanda umsebenzi wee-enzyme ezintathu kunye nokukhusela i-RNA ekuthotyweni.Nangona kunjalo, ayisebenzi ngokuchasene ne-RNase 1, i-RNase T1, i-S1 Nuclease, i-RNase H okanye i-RNase esuka kwi-Aspergillus.I-Murine RNase inhibitor yavavanywa yi-RT-PCR, RT-qPCR kunye ne-IVT mRNA, kwaye yayihambelana neentlobo ezahlukeneyo zentengiso ye-Reverse transcriptases, i-DNA polymerases kunye ne-RNA polymerases.

Xa kuthelekiswa ne-inhibitors ye-RNase yabantu, i-murine RNase inhibitor ayinayo i-cysteines ezimbini ezithintekayo kakhulu kwi-oxidation ebangela ukungasebenzi kwe-inhibitor.Oko kwenza ukuba izinze kwiindawo eziphantsi ze-DTT (ngaphantsi kwe-1 mM).Olu phawu luyenza ilungele ukusetyenziswa kwiimpendulo apho ugxininiso oluphezulu lwe-DTT luchasene nokusabela (umz. Ixesha lokwenyani i-RT-PCR).

Ngaphambili: I-Hotstart Taq DNA Polymerase HC1012A

Okulandelayo: I-Hs Taq DNA Polymerase(iGlycerol-free)

Aisicelo

Le mveliso inokusetyenziswa ngokubanzi kulo naluphi na uvavanyo apho uphazamiseko lwe-RNase lunokwenzeka ukunqanda ukuthotywa kwe-RNA, efana nale:

1.I-synthesis ye-strand yokuqala ye-cDNA, i-RT-PCR, i-RT-qPCR, njl.

2.Ikhusela i-RNA ekuthotyweni ngexesha loshicilelo/uguqulelo lwe-in vitro (umzekelo, inkqubo yophindaphindo lwentsholongwane kwi-vitro).

3.Ukuvinjelwa komsebenzi we-RNase ngexesha lokuhlukana kwe-RNA kunye nokuhlanjululwa.

Iimeko zokuGcina

Imveliso inokugcinwa kwi -25～- 15 ℃, isebenza iminyaka emi-2.

Isithinteli sogcino

50 mM KCl, 20 mM HEPES-KOH (pH 7.6, 25 ℃), 8 mM DTT kunye ne-50% glycerol.

Inkcazo yeyunithi

Isixa se-murine RNase inhibitor efunekayo ukuvimbela umsebenzi we-5ng ye-ribonuclease A nge-50% ichazwe njengeyunithi enye (U).

Ubunzima bemolekyuli yeprotheni

Ubunzima be-molecular of murine RNase inhibitor yi-50 kDa.

Ulawulo lwemeko

I-Exonuclease Umsebenzi:

I-40 U ye-murine RNase inhibitor kunye ne-1 μg λ -Hind III digest DNA kwi-37℃ kwiiyure ze-16 ayivelisi ukuthotywa njengoko kunqunywe yi-agarose gel electrophoresis.
Umsebenzi we-Endonuclease:

I-40 U ye-murine RNase inhibitor kunye ne-1μ g λ DNA kwi-37℃ kwiiyure ze-16 ayivelisi ukuthotywa njengoko kunqunywe yi-agarose gel electrophoresis.

Ukuqhula Umsebenzi:

I-40U ye-murine RNase inhibitor ene-1μ g pBR322 kwi-37℃ kwiiyure ze-16 ivelisa akukho ukuthotywa njengoko kunqunywe yi-agarose gel electrophoresis.

RNase Umsebenzi:

I-40U ye-murine RNase inhibitor kunye ne-1.6μ g MS2 RNA ye-4h kwi-37℃ ayivelisi ukuthotywa njengoko kunqunywe yi-agarose gel electrophoresis.

E.coli DNA:

I-40 U ye-murine RNase inhibitor ihlolwe ubukho be-E. coli genomic DNA usebenzisa i-TaqMan qPCR kunye ne-primers ekhethekileyo ye-E. coli 16S rRNA locus.Ungcoliseko lwe-E. coli genomic DNA ngu ≤ 0.1 pg/40 U.

Notes

I-1.I-oscillation enobundlobongela okanye ukuvuselela kuya kubangela ukungasebenzi kwe-enzyme.

2.Olona luhlu lweqondo lokushisa oluphezulu lwale inhibitor lwaluyi-25-55 ℃, kwaye lwalungasebenzi kwi-65℃ nangaphezulu.

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

I-4.I-inhibition yomsebenzi we-RNase ifunyenwe kuluhlu olubanzi lwe-pH (i-pH 5-9 yonke yayisebenza), kwaye umsebenzi ophezulu wabonwa kwi-pH 7-8.

5. Ekubeni ii-ribonucleases zihlala zigcina umsebenzi phantsi kweemeko ze-denaturing, kufuneka kuthatyathwe ukhathalelo ukuze kugwenywe ukwenza iimolekyuli ze-RNase Inhibitor eziye zadibana ne-ribonuclease.Ukuthintela ukukhutshwa kwe-ribonuclease esebenzayo, amaqondo obushushu angaphezu kwama-50 °C kunye noxinzelelo oluphezulu lwe-urea okanye ezinye ii-denaturing agents kufuneka ziphetshwe.