UDG/UNG Enzymes
Hiki i ka UDG (uracil DNA glycosylase) ke hoʻopale i ka hydrolysis o ka loulou N-glycosidic ma waena o ke kumu uracil a me ka iwi kuamoʻo kō-phosphate ma ssDNA a me dsDNA.Hiki iā ia ke hoʻomalu i ka pollution aerosol a kūpono i nā ʻōnaehana biology molecular maʻamau e like me PCR, qPCR, RT-qPCR a me LAMP.
Nā kikoʻī
| Hoʻokipa Hōʻike | Recombinant E. coliwith uracil DNA glycosylase gene |
| Kaumaha Molekala | 24. 8kDa |
| Maemae | ≥95% (SDS-PAGE) |
| ʻAʻohe wela | 95 ℃, 5-10 min |
| Wehewehe Unite | Hoʻokahi ʻāpana (U) i wehewehe ʻia ʻo ia ka nui o ka enzyme e koi ʻia e hoʻopololei i ka hydrolysis o l μg dU i loaʻa i ka dsDNA i 30 mau minuke ma 25 ℃. |
Waihona
Pono e mālama ʻia ka huahana ma-25 ℃ ~ -15 ° C no ʻelua makahiki.
Nā kuhikuhi
1.Ka hoʻomākaukau ʻana i ka hui ʻana o ka PCR e like me ka ʻōnaehana hahai
| Nā ʻāpana | Volume (μL) | Hoʻoholo hope |
| 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 | - |
| Kumu1 (10μmol/L) | 2 | 0.4 μmol/L |
| Kumuhana 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 | A hiki i 50 | - |
Nānā: Wahi a nā koi hoʻokolohua, hiki ke hoʻololi ʻia ka ʻike hope loa o ka dUTP ma waena o 0.2-0.6 mmol/L, a hiki ke hoʻohui ʻia ka 0.2 mmol/L dTTP.
2.Kaʻina hana hoʻonui
| Kaʻina kaʻapuni | Mahana | Manawa | Nā pōʻaiapuni |
| ka hoʻohaʻahaʻa ʻana i ka laʻana me dU | 25 ℃ | 10 min | 1 |
| Hoʻoulu ʻia ʻo UDG, denaturation mua | 95 ℃ | 5 ~ 10 mau minuke | 1 |
| Denaturation | 95 ℃ | 10 sec |
30-35 |
| Hoʻopili | 60 ℃ | 20 sec | |
| Hoʻonui | 72 ℃ | 30 sec/kb | |
| Hoʻonui hope | 72 ℃ | 5 min | 1 |
Nānā: Hiki ke hoʻololi i ka manawa pane ma 25 ° C i loko o 5-10mins e like me nā koi hoʻokolohua.
Nā memo
1.Hoʻoikaika ʻo UDG i ka hapa nui o ka PCR.
2.Pono e mālama ʻia nā enzyme i loko o ka pahu hau a i ʻole ma ka ʻauʻau hau ke hoʻohana ʻia, a pono e mālama ʻia ma-20°C ma hope koke o ka hoʻohana ʻana.
3.E ʻoluʻolu e ʻaʻahu i ka PPE pono, e like me ka lole lab a me nā mīkina lima, e hōʻoia i kou olakino a palekana!
