Rnase Inhibitor (Glycerol bilaash ah)
Murine RNase inhibitor waa xuubka RNase dib-u-dhaqaale oo la muujiyey oo laga nadiifiyey E.coli.Waxay ku xidhan tahay RNase A, B ama C ee saamiga 1: 1 iyada oo loo marayo isku xidhid aan wadajir ahayn, taas oo ka hortagaysa dhaqdhaqaaqa saddexda enzymes iyo ka ilaalinta RNA ee hoos u dhaca.Si kastaba ha ahaatee, waxtar uma laha RNase 1, RNase T1, S1 Nuclease, RNase H ama RNase ka Aspergillus.Murine RNase inhibitor waxaa tijaabiyay RT-PCR, RT-qPCR iyo IVT mRNA, waxayna la jaanqaadi jirtay qoraalada kala duwan ee ganacsiga ee Reverse transcriptases, polymerases DNA iyo polymerases RNA.
Marka la barbardhigo bini'aadamka RNase inhibitors, murine RNase inhibitor kuma jiraan laba cysteinees kuwaas oo aad ugu nugul oksaydheynta taas oo keenta firfircoonida xakameynta.Taas oo ka dhigaysa mid xasiloon marka la eego xaddi yar oo DTT ah (in ka yar 1 mM).Habkani waxa uu ka dhigayaa mid ku haboon in loo isticmaalo falcelinta halka xaddi badan oo DTT ah ay lid ku tahay falcelinta (tusaale-waqtiga dhabta ah ee RT-PCR).
Acodsi
Alaabtan waxa si weyn loogu isticmaali karaa tijaabo kasta oo faragelinta RNase ay suurtogal tahay si looga fogaado xaalufka RNA, sida:
1.First-strand cDNA synthesis, RT-PCR, RT-qPCR, iwm;
2.Ka ilaali RNA hoos-u-dhac ku yimaadda qoraalka/tarjumaada in-vitro (sida fayraska ku-noqoshada gudaha vitro);
3. Joojinta dhaqdhaqaaqa RNase inta lagu jiro go'doominta RNA iyo nadiifinta.
Xaaladaha Kaydinta
Ku kaydi -25～-15℃;
Wareegyada dhalaalay ≤ 5 jeer;
Shaqeynaya 1 sano
Qeexitaanka unugga
Hal unug ayaa lagu qeexay cadadka RNase Inhibitor ee looga baahan yahay inuu joojiyo dhaqdhaqaaqa 5 ng ee RNase A 50%.
Miisaanka molecular
RNase Inhibitor (Glycerol-free) waa borotiin 50 kDa ah.
Xakamaynta tayada
Exonuclease Hawsha:
Ku-soo-baxa 40 U ee enzyme leh 1 μg λ-Hind III dheefshiidka DNA-da saacadaha 16 ee 37 ° C waxay keentay in aan la ogaan karin hoos u dhac ku yimaada DNA-da sida lagu go'aamiyo gel electrophoresis.
Waxqabadka Endonuclease:
Ku-soo-baxa 40 U of enzyme leh 1 μg λ DNA 16 saacadood oo ah 37 ° C waxay keentay in aan la ogaan karin hoos u dhac ku yimaada DNA-da sida lagu go'aamiyo jel electrophoresis.
Naajin Hawsha:
Ku-soo-baxa 40 U of enzyme leh 1 μg pBR322 muddo 16 saacadood ah 37 ℃ waxay keentay in aan la ogaan karin hoos u dhac ku yimaada DNA-da sida lagu go'aamiyay jel electrophoresis.
RNase Hawsha:
Ku-soo-baxa 40 U of enzyme leh 1.6 μg MS2 RNA muddo 4 saacadood ah 37 ℃ waxay keentay in aan la ogaan karin hoos u dhac RNA ah sida lagu go'aamiyay jel electrophoresis.
E.coli DNA:
40 U of Enzyme waxaa lagu ogaadaa TaqMan qPCR.DNA-da E.coli waa ≤ 0. 1pg/40U.
Notes
1. Ha ruxin ama si xoog leh ha u kicin si aad uga hortagto firfircoonida enzyme.
2.RNase inhibitor waxa uu ku shaqeeyaa heerkul u dhexeeya 25℃ ilaa 55℃ waxana aanu shaqaynayn ≥65℃
3. Lama joojinayo dhaqdhaqaaqa RNase H, RNase 1, iyo RNase T1.
4. Kala duwanaanshaha pH ee lagu joojinayo dhaqdhaqaaqa RNase waa mid ballaaran (firfircoon pH 5-9), oo muujinaya dhaqdhaqaaqa ugu badan ee pH 7-8.
