UDG/UNG Enzymes
UDG (uracil DNA glycosylase) na iya haifar da haɓakar hydrolysis na haɗin N-glycosidic tsakanin tushen uracil da kashin baya na sukari-phosphate a cikin ssDNA da dsDNA.Yana iya sarrafa gurɓacewar iska cikin sauƙi kuma ya dace da tsarin ilimin halitta gama gari kamar PCR, qPCR, RT-qPCR da LAMP.
Ƙayyadaddun bayanai
| Mai watsa shiri Magana | Recombinant E. coli tare da uracil DNA glycosylase gene |
| Nauyin Kwayoyin Halitta | 24.8k da |
| Tsafta | ≥95% (SDS-PAGE) |
| Rashin kunna zafi | 95 ℃, 5 ~ 10 min |
| Ma'anar Naúrar | An ayyana raka'a ɗaya (U) azaman adadin enzyme wanda ake buƙata don haɓaka hydrolysis na l μg dU mai ɗauke da dsDNA a cikin mintuna 30 a 25℃. |
Adana
Ya kamata a adana samfurin a -25 ℃ ~ -15 ° C na shekaru biyu.
Umarni
1.Shiri na cakuda amsawar PCR bisa ga tsarin mai zuwa
| Abubuwan da aka gyara | girma (μL) | Natsuwa na ƙarshe |
| 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 |
| Samfurin DNA | X | - |
| Farko 1 (10μmol/L) | 2 | 0.4 μmol/L |
| Farko 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 | Har zuwa 50 | - |
Lura: Dangane da buƙatun gwaji, za a iya daidaita ƙaddamarwar ƙarshe na dUTP tsakanin 0.2-0.6 mmol/L, kuma 0.2 mmol/L dTTP za a iya ƙarawa da zaɓi.
2.Hanyar haɓakawa
| Zagayowar mataki | Zazzabi | Lokaci | Zagaye |
| dU mai ƙunshe da lalatar samfuri | 25 ℃ | Minti 10 | 1 |
| Kunna UDG, samfuri na farko denaturation | 95 ℃ | 5 ~ 10 min | 1 |
| Denaturation | 95 ℃ | 10 dakika |
30-35 |
| Annealing | 60 ℃ | dakika 20 | |
| Tsawaita | 72 ℃ | 30 seconds/kb | |
| Ƙarshe na ƙarshe | 72 ℃ | 5 min | 1 |
Lura: Ana iya daidaita lokacin amsawa a 25 ° C a cikin 5-10mins bisa ga buƙatun gwaji.
Bayanan kula
1.UDG yana aiki a mafi yawan abubuwan da ake buƙatar amsawa na PCR.
2.Ya kamata a adana enzymes a cikin akwatin kankara ko kuma a kan wanka na kankara lokacin amfani da shi, kuma ya kamata a adana shi a-20 ° C nan da nan bayan amfani.
3.Da fatan za a saka PPE da ake buƙata, irin wannan rigar lab da safar hannu, don tabbatar da lafiyar ku da amincin ku!
