Rnase A
Ribonuclease A(RNaseA) waa polypeptide-ka-xidhan oo ka kooban 4 curaarta disulfide oo miisaankeedu yahay 13.7 kDa.RNase Ais waa endoribonuclease kaas oo si gaar ah hoos u dhigaya RNA hal-xadhigleed ee C iyo U hadhaaga.Gaar ahaan, kala-goynta waxay aqoonsanaysaa isku-xidhka fosfodiester-ka ee ay sameeyeen 5'-ribose ee nucleotide iyo kooxda fosfateetka ee 3'-ribose ee nucleotide pyrimidine ee ku xiga, si fosfatiyeyaasha 2,3'-Cyclic lagu hydrolyzed 3 u dhiganta. phosphates nucleoside (tusaale, pG-pG-pC-pA-pG waxaa kala jeexjeexay RNase A si loo dhaliyo pG-pG-pCp iyo A-PG).RNase A waa kan ugu firfircoon ee kala jeexjeexa RNA-geesoodka ah.Xoog saarista shaqada ee lagu taliyay waa 1-100 μ G/mL, oo la jaan qaadaya hababka falcelinta ee kala duwan.Isku-duubnaanta milixdu yar tahay (0-100 mM NaCl) ayaa loo isticmaali karaa in lagu gooyo RNA hal-xadhigle ah, RNA laba-geesood ah, iyo RNA silsilado ay sameeyeen isku-dhafka RNA-DNA.
Si kastaba ha noqotee, milix badan (≥0.3 M), RNase A kaliya waxay si gaar ah u kala dillaacdaa RNA hal-xadhigle ah.
RNase A waxaa inta badan loo isticmaalaa in meesha laga saaro RNA inta lagu jiro diyaarinta DNA-da plasmid ama genomic DNA.Haddii DNAse firfircoon tahay iyo in kale inta lagu jiro habka diyaarinta waxay si fudud u saameyn kartaa falcelinta.Habka dhaqameed ee lagu kariyo biyaha qubeyska ayaa loo isticmaali karaa in la joojiyo dhaqdhaqaaqa DNA-ga.Alaabtani kuma jiraan DNA-ga iyo borotiinka, umana baahna daawaynta kulaylka ka hor inta aan la isticmaalin.Intaa waxaa dheer, badeecadan sidoo kale waxaa loo isticmaali karaa tijaabooyinka bayoloji molecular sida falanqaynta ilaalinta RNase iyo falanqaynta taxanaha RNA.
Xaaladaha kaydinta
Alaabta waa in lagu kaydiyaa at -25℃~-15℃ 2 sano.
Tilmaamaha
| Muuqashada | Budada |
| Tirada | 100mg / 1g |
| Nooca alaabta | RNase A |
Tilmaamaha
Tani waa mid ka mid ah hababka caadiga ah ee diyaarinta RNase A xal kaydinta.Sidoo kale waa la diyaarin karaahabab kale oo waafaqsan hababka dhaqanka ee shaybaarka ama suugaanta tixraaca (sidasi toos ah ugu milma 10 mM Tris-HCl, pH 7.5 ama Tris-NaCl
1. Isticmaal 10 mM sodium acetate (pH 5.2) si aad u diyaariso 10 mg/mL ee RNase A xal kaydinta.
2. Kuleylinta 100 ℃ 15 daqiiqo.
3. Ku qabow heerkulka qolka, ku dar 1/10 mug 1 M Tris-HCl (pH 7.4), ku hagaaji pH ilaa 7.4tusaale ahaan, ku dar 500 ml oo ah 10 mg/mL RNase xal kaydinta 1 M Tris-HCl, pH7.4).
4. Baakad hoose oo ah -20℃ kaydinta barafaysan, taas oo noqon karta mid xasilloon ilaa 2 sano.
[Xusuusin]:
Marka lagu karkariyo xalalka RNaseA ee xaaladaha dhexdhexaadka ah, roobab RNase ayaa sameyn doona;Ku kari pH hoose, iyo haddii ay jirto roobab, waa la arki karaa, taas oo ay sababi karto joogitaanka borotiinka wasakhaysan.Haddii wasakhda la helo ka dib karkarinta, wasakhda waxaa lagu saari karaa xawaaraha sare ee centrifugation (13000rpm), ka dibna si hoose loogu shubo kaydinta qaboojinta.
Xusuusin
Fadlan xidho PPE-ga lagama maarmaanka ah, sida jaakada shaybaadhka iyo galoofyada, si loo hubiyo caafimaadkaaga iyo badbaadadaada.
