Ultra Nuclease GMP-grade
Bisadda No: HC2016A
UltraNuclease GMP-grade waxa lagu muujiyay oo lagu nadiifiyaa Escherichia coli (E.coli) iyada oo la farsameeyay hidde ahaan loona diyaariyay agagaaraha GMP.Waxay yarayn kartaa viscosity ee supernatant unug iyo lysate unug ee cilmi-baarista sayniska, kordhinta hufnaanta borotiinka iyo kor u qaadida cilmi-baarista functional protein.Alaabtu waxay sidoo kale hoos u dhigi kartaa hadhaaga aashitada nucleic-ka ah ee martida loo yahay pg-grade, hagaajinta waxqabadka iyo badbaadada alaabada bayoolojiga ee codsiyada ay ku jiraan nadiifinta fayraska, soo saarista tallaalka, iyo borotiinka/polysaccharide soosaarka dawooyinka.Ka sokow, sheyga ayaa sidoo kale lagu dabaqi karaa si looga hortago isku dhafka unugyada mononuclear dhiigga ee bini'aadamka (PBMC) ee daaweynta unugyada iyo horumarinta tallaalka.
UltraNuclease waxaa lagu bixiyaa qaab reagent jeermis dilay, oo laga soo saaray kayd (20mM Tris-HCL pH 8.0, 2mM MgCl, 20 mM NaCl, 50% glycerin), oo leh muuqaalka dareere aan midab lahayn, oo hufan.Alaabtan waxaa soo saaray shuruudaha nidaamka GMP waxaana lagu bixiyaa qaab dareere ah.
Qaybaha
UltraNuclease GMP-darajada (250 U/μL)
Xaaladaha Kaydinta
Alaabta waxaa lagu raray baraf qallalan waxaana lagu kaydin karaa -25℃~-15°C muddo laba sano ah.
Haddii badeecada la furay oo lagu kaydiyay 4℃ in ka badan toddobaad, waxaan kugula talineynaa shaandhayntaalaabta si looga hortago wasakhowga microbial.
Tilmaamaha
| Martigaliyaha Hadalka | Dib-u-habaynta E. coli oo leh hidda-wadaha UltraNuclease |
| Miisaanka Molecular | 26.5kDa |
| barta soelectric | 6.85 |
| daahirnimo | ≥99% (SDS-PAGE) |
| Kaydinta kaydinta | 20mM Tris-HCL pH 8.0, 2mM MgCl, 20mM NaCl, 50% glycerin |
|
Qeexitaan Unug | Qeexida hal unug Waxqabad (U) waa qadarka ensaymka loo isticmaalobeddel qiimaha nuugista ΔA260 1.0 30 daqiiqo gudahood 2. 625 mLhabka falcelinta ee 37 ℃ oo leh pH 8.0 (oo u dhiganta dhammaystirka dheefshiidka37 μg salmon oo shahwada DNA galay oligonucleotides). |
Tilmaamaha
1. Tusaale Ururinta
Unugyada ku dheggan: ka saar dhexdhexaadka, ku maydh unugyada PBS, oo ka saar kuwa sarreeya.
Unugyada ka joojinta: ku soo ururi unugyada iyagoo centrifugation ah, ku dhaq unugyada PBS, centrifuge at 6,000rpm ilaa 10 daqiiqo, qaado pellet.
Escherichia coli: Bakteeriyada ku soo qaado centrifugation, hal mar ku dhaq PBS, centrifuge at 8,000rpm ilaa 5 daqiiqo, oo qaado pellet-ka.
2. Tusaalaha Daawaynta
Ku dawee pellets unug la ururiyey adigoo isticmaalaya lysis baffer marka loo eego cufka(g) ilaa mugga(mL)1:(10-20), ama habab farsamo ama kiimiko oo baraf ah ama heerkulka qolka
109 unug).
3. Daaweynta Enzyme
Ku dar 1-5mM MgCl habka falcelinta oo ku hagaaji pH ilaa 8-9.
Ku dar UltraNuclease iyadoo loo eegayo saamiga 250 Cutub si aad u shiiddo 1 g oo ah pellets unug, ku dhex jir 37℃ in ka badan 30 daqiiqo.Fadlan tixraac foomka "Waqtiga falcelinta lagu taliyay" si aad u dooratomuddada daaweynta.
4. Sareeye Ururinta
Centrifuge at 12,000 rpm 30 daqiiqo oo soo qaado heerka sare.
Fiiro gaar ah: Haddii xalku yahay acidic ama alkaline, ama ay ku jiraan xaddi badan oo milix ah, saabuunta, amaDenaturants, fadlan kordhi qiyaasta enzyme ama u kordhi wakhtiga daaweynta si waafaqsan.
Xaalad falcelin lagu taliyayons
| Halbeegga | Xaaladda ugu Fiican | Xaaladda Waxtarka leh |
| Mg²+Diirada | 1-5 mm | 1-10 mm |
| pH | 8-9 | 6-10 |
| Heerkulka | 37℃ | 0-42℃ |
| Isku-duubnida DTT | 0-100 mm | >0 mM |
| Isku-darka Mercaptoethanol | 0-100 mm | >0 mM |
| Isku-xidhka Monovalent Cation | 0-20 mm | 0-150 mm |
| Fosfate lon Diiradda | 0-10 mm | 0-100 mm |
lagu taliyay Falcelinta Waqtiga (37 ℃, 2 mM Mg²+, pH 8.0)
| Qadarka UltraNuclease (U-fiirsashada u dambaysa) | Waqtiga Falcelinta |
| 0.25 U/ml | >10 saac |
| 2.5 U/ml | >4saac |
| 25 U/ml | 30 daqiiqo |
Xusuusin:
Fadlan xidho PPE-ga lagama maarmaanka ah, sida jaakada shaybaadhka iyo galoofyada, si loo hubiyo caafimaadkaaga iyo badbaadadaada!
