Rnasa A
Ribonuclease A(RNaseA) polypeptide ne mai madauri guda ɗaya mai ɗauke da 4 disulfide bonds tare da nauyin kwayoyin halitta na kusan 13.7 kDa.RNase Ai shi ne endoribonuclease wanda ke lalata RNA mai madauri ɗaya musamman a ragowar C da U.Musamman, cleavage ya gane haɗin phosphodiester da aka kafa ta hanyar 5'-ribose na nucleotide da ƙungiyar phosphate akan 3'-ribose na pyrimidine nucleotide na kusa, don haka 2,3'-Cyclic phosphates suna hydrolyzed zuwa daidai 3. 'Nucleoside phosphates (misali, pG-pG-pC-pA-pG RNase A ce ta ware don samar da pG-pG-pCp da A-PG).RNase A shine mafi aiki a cikin tsage RNA mai madauri ɗaya.Ƙaddamar da aikin da aka ba da shawarar shine 1-100 μ G / ml, mai jituwa tare da tsarin amsawa daban-daban.Za'a iya amfani da ƙarancin ƙarancin gishiri (0-100 mM NaCl) don yanke RNA mai ɗauri ɗaya, RNA mai ɗauri biyu, da sarƙoƙin RNA waɗanda aka kafa ta hanyar haɓakar RNA-DNA.
Koyaya, a babban maida hankali na gishiri (≥0.3 M), RNase A kawai yana tsinke RNA mai madauri ɗaya.
An fi amfani da RNase A don cire RNA yayin shirye-shiryen DNA na plasmid ko DNA na genomic.Ko DNAse yana aiki ko a'a yayin tsarin shirye-shiryen zai iya tasiri cikin sauƙi.Ana iya amfani da hanyar gargajiya ta tafasa a cikin ruwan wanka don hana ayyukan DNAse.Wannan samfurin baya ƙunshi DNAase da protease, kuma baya buƙatar maganin zafi kafin amfani.Bugu da ƙari, ana iya amfani da wannan samfurin a cikin gwaje-gwajen ilimin halitta kamar nazarin kariyar RNase da nazarin jerin RNA.
Yanayin ajiya
Ya kamata a adana samfurin a -25 ℃ ~ - 15 ℃ na shekaru 2.
Ƙayyadaddun bayanai
| Bayyanar | Foda |
| Yawan | 100mg/1g |
| Nau'in samfur | RNase A |
Umarni
Wannan shine ɗayan hanyoyin gama gari don shirya mafita na RNase A.Hakanan za'a iya shirya shita wasu hanyoyin bisa ga hanyoyin gargajiya a cikin dakin gwaje-gwaje ko wallafe-wallafe (kamarNarkar da kai tsaye a cikin 10 mM Tris-HCl, pH 7.5 ko maganin Tris-NaCl)
1. Yi amfani da 10 mM sodium acetate (pH 5.2) don shirya 10 mg / ml na RNase A ajiya bayani.
2. Dumama a 100 ℃ na 15 min.
3. Cool zuwa zafin jiki, ƙara ƙarar 1/10 na 1 M Tris-HCl (pH 7.4), daidaita pH zuwa 7.4 (donmisali, ƙara 500 ml na 10 mg/mL RNase maganin ajiya 1 M Tris-HCl, pH7.4).
4. Sub-cushe a -20 ℃ don daskararre ajiya, wanda zai iya zama barga har zuwa shekaru 2.
[Bayanai]:
Lokacin tafasa maganin RNaseA a ƙarƙashin yanayin tsaka tsaki, hazo na RNase zai haifar;Tafasa shi a ƙasan pH, kuma idan akwai hazo, ana iya lura da shi, wanda zai iya haifar da kasancewar gurɓataccen furotin.Idan aka sami laka bayan tafasa, za'a iya cire ƙazanta ta hanyar centrifugation mai sauri (13000rpm), sannan a yi ƙasa da ƙasa don ajiya mai daskarewa.
Bayanan kula
Da fatan za a saka PPE da ake buƙata, irin wannan rigar lab da safar hannu, don tabbatar da lafiyar ku da amincin ku.
