UDG/UNG Enzymes
UDG (uracil DNA glycosylase) inogona kukonzera hydrolysis yeN-glycosidic link pakati pe uracil base uye shuga-phosphate musana mu ssDNA uye dsDNA.Inogona kudzora nyore kusvibiswa kweaerosol uye inokodzera zvakajairika mamorekuru biology masisitimu akadai sePCR, qPCR, RT-qPCR uye LAMP.
Zvinotsanangurwa
| Expression Host | Recombinant E. coliwith uracil DNA glycosylase gene |
| Molecular Weight | 24. 8kDa |
| Kuchena | ≥95% (SDS-PEJI) |
| Heat Inactivation | 95 ℃, 5 ~ 10 min |
| Chikamu Tsanangudzo | Imwe unit (U) inotsanangurwa sehuwandu hwe enzyme yaidiwa kukonzeresa hydrolysis yel μg dU-ine dsDNA mumaminitsi makumi matatu pa25 ℃. |
Storage
Chigadzirwa chacho chinofanira kuchengetwa pa-25 ℃ ~ -15 ° C kwemakore maviri.
Mirayiridzo
1.Kugadzirira kwePCR reaction musanganiswa zvinoenderana nekutevera system
| Zvikamu | Vhoriyamu (μL) | Final concentration |
| 10×PCR Buffer (Mg²+Plus) | 5 | 1× |
| 25 mmol/LMgCl | 3 | 1.5 mmol/L |
| dUTP (10 mmol/L) | 3 | 0.6 mmol/L |
| dCTP/dGTP/dATP/dTTP(10mmol/Leach) | 1 | 0.2 mmol / Leach |
| DNA template | X | - |
| Primer1 (10μmol/L) | 2 | 0.4 μmol/L |
| Primer 2 (10μmol/L) | 2 | 0.4 μmol/L |
| Taq DNA Polymerase (5 U/μL) | 0. 5 | 0. 05 U/μL |
| Uracil DNA Glycosylase (UDG/UNG), 1 U/μL | 1 | 1 U/μL |
| ddH₂O | Kusvikira ku50 | - |
Cherechedza: Zvinoenderana nezvinodiwa zvekuedza, iyo yekupedzisira kusungirirwa kwedUTP inogona kugadziriswa pakati pe0.2-0.6 mmol / L, uye 0.2 mmol / L dTTP inogona kuwedzerwa zvakasarudzwa.
2.Amplification maitiro
| Cycle step | Tembiricha | Nguva | Cycles |
| dU-ine template degradation | 25℃ | 10mins | 1 |
| UDG activation, template yekutanga denaturation | 95℃ | 5 ~ 10mins | 1 |
| Denaturation | 95℃ | 10 sec |
30-35 |
| Annealing | 60 ℃ | 20 sec | |
| Extension | 72℃ | 30sec/kb | |
| Final extension | 72℃ | 5 min | 1 |
Cherechedza: Nguva yekuita pa25 ° C inogona kugadziriswa mukati me5-10mins maererano nekuedza zvinodiwa.
Notes
1.UDG inoshanda mune dzakawanda PCR reaction buffers.
2.Ma Enzymes anofanirwa kuchengetwa mubhokisi rechando kana pabhavhu rechando kana ashandiswa, uye anofanira kuchengetwa pa-20 ° C pakarepo mushure mekushandisa.
3.Ndokumbira upfeke PPE inodiwa, jasi rakadaro uye magurovhosi, kuve nechokwadi chehutano hwako uye kuchengetedzeka!
