Rnase Inhibitor (Glycerol kyauta)
Murine RNase inhibitor shine mai hana murine RNase wanda aka bayyana kuma an tsarkake shi daga E.coli.Yana ɗaure zuwa RNase A, B ko C a cikin rabo na 1:1 ta hanyar haɗin kai mara daidaituwa, ta haka yana hana ayyukan enzymes uku da kare RNA daga lalacewa.Koyaya, Ba shi da tasiri akan RNase 1, RNase T1, S1 Nuclease, RNase H ko RNase daga Aspergillus.Murine RNase inhibitor an gwada ta RT-PCR, RT-qPCR da IVT mRNA, kuma ya dace da nau'ikan bayanan Reverse na kasuwanci, polymerases na DNA da RNA polymerases.
Idan aka kwatanta da masu hana RNase na ɗan adam, mai hana RNase na murine ba ya ƙunshi nau'in cysteines guda biyu waɗanda ke da matukar damuwa ga oxidation wanda ke haifar da rashin kunna mai hanawa.Wannan yana mai da shi kwanciyar hankali a ƙananan adadin DTT (kasa da 1 mM).Wannan fasalin ya sa ya dace don amfani a cikin halayen inda babban taro na DTT ke da illa ga abin da ya faru (misali RT-PCR na ainihi).
Aaikace-aikace
Ana iya amfani da wannan samfur ko'ina a kowane gwaji inda tsangwama na RNase zai yiwu don guje wa lalatawar RNA, kamar:
1.First-strand cDNA kira, RT-PCR, RT-qPCR, da dai sauransu;
2.Kare RNA daga lalacewa a cikin fassarar in-vitro / fassarar (misali kwafi na hoto a cikin vitro);
3.Hana ayyukan RNase yayin keɓewar RNA da tsarkakewa.
Yanayin Ajiya
Adana a -25～-15 ℃;
Daskare-narke hawan keke ≤ sau 5;
Yana aiki na shekaru 1.
Ma'anar raka'a
An bayyana raka'a ɗaya azaman adadin RNase Inhibitor da ake buƙata don hana ayyukan 5 ng na RNase A da 50%.
Nauyin kwayoyin halitta
RNase Inhibitor (Glycerol-free) furotin ne na kDa 50.
Kula da inganci
Exonuclease Ayyuka:
Ƙaddamar da 40 U na enzyme tare da 1 μg λ-Hind III narkar da DNA na 16 hours a 37 ° C ya haifar da rashin lahani na DNA kamar yadda gel electrophoresis ya ƙaddara.
Ayyukan Endonuclease:
Ƙaddamar da 40 U na enzyme tare da 1 μg λ DNA na tsawon sa'o'i 16 a 37 ° C ya haifar da rashin lahani na DNA kamar yadda gel electrophoresis ya ƙaddara.
Yin waƙa Ayyuka:
Ƙaddamar da 40 U na enzyme tare da 1 μg pBR322 na tsawon sa'o'i 16 a 37 ℃ ya haifar da rashin lahani na DNA kamar yadda gel electrophoresis ya ƙaddara.
RNase Ayyuka:
Shigar da 40 U na enzyme tare da 1.6 μg MS2 RNA na tsawon awanni 4 a 37 ℃ ya haifar da rashin lahani na RNA kamar yadda gel electrophoresis ya ƙaddara.
E.coli DNA:
An gano 40 U na Enzyme ta TaqMan qPCR.E.coli DNA shine ≤ 0. 1pg/40U.
Notes
1.Kada a girgiza ko motsawa da ƙarfi don hana rashin kunna enzyme.
2.RNase inhibitor yana aiki a yanayin zafi daga 25 ℃ zuwa 55 ℃ kuma an kunna shi a ≥65 ℃.
3.Ba a hana ayyukan RNase H, RNase 1, da RNase T1 ba.
4.The pH kewayon don hana ayyukan RNase yana da fadi (aiki a pH 5-9), yana nuna matsakaicin aiki a pH 7-8.
