Tembiricha Inonzwa UNG
Temperature Sensitive UNG (TS-UNG) inowanikwa nerecombinant expression in E. coli.Iyo enzyme inokonzeresa kuburitswa kwemahara uracil kubva muuracil-ine imwe- uye ine mbiri-yakasungwa DNA uye haishande kurwisa RNA.Kuenzaniswa neyakajairwa UNG enzyme yeE. coli gene mabviro, TS-UNG enzyme ine chiitiko chepamusoro pakudziya kwakaderera (20 ℃ ~ 37 ℃) uye inonzwa tembiricha uye inovharika nyore (50 ℃), ichidzivirira kuderedzwa kweDUTP-ine amplification. zvigadzirwa patembiricha yekamuri neyakasara chiitiko chinogona kusara mushure mekusashanda kweyakajairwa UNG enzyme.Naizvozvo, TS-UNG enzyme haina kungokodzera PCR kusvibiswa kwekudzivirira maitiro, asi zvakare inoenderana zvakanaka neRT-PCR yekusimudzira chirongwa uye inogona kushandiswa muRT-PCR yekudzivirira kusvibiswa maitiro.
Recommended Application
Contamination Prevention Amplification
Storage Condition
-20°C kuitira kuchengetedza kwenguva refu, inofanira kusanganiswa zvakanaka isati yashandiswa, dzivisa kugaro chando-nyorova.
Storage buffer
20 mM Tris-HCl (pH 7.5) , 100 mM NaCl, 0.1 mM EDTA, 1 mM DTT, Stabilizer, 50% Glycerol.
Chikamu Tsanangudzo
Kuwanda kweenzyme inodiwa kuderedza 1µg yeDNA imwe chete ine mabhesi eDU muawa imwe pa37 ° C i1 unit yebasa (U).
Quality Control
1.SDS-PAGE electrophoretic kuchena kukuru kupfuura 98%
2.Kuzvidzisa basa, batch-to-batch control, kugadzikana
3.Hapana exogenous nuclease chiitiko, hapana exogenous endonuclease kana exonuclease kusvibiswa.
Mirayiridzo
| Zvikamu | Vhoriyamu (μL) | Final concentration |
| 10 × PCR Buffer (dNTP yemahara, Mg²+mahara) | 5 | 1× |
| dUTPs (dCTP, dGTP, dATP) | - | 200 μM |
| dUTP (tsiva dTTP) | - | 200-600 μM |
| 25 mM MgCl2 | 2-8 μL | 1-4 mM |
| 5 U/μL Taq | 0.25 | 1.25 U |
| 1 U/μLTS-UNG | 0.5 (0.1-0.5) | 0.5 U (0.1-0.5U) |
| 25 × Primer Mixa | 2 | 1× |
| Template | - | ＜1μg/maitiro |
| ddH₂O | kusvika ku50 | - |
Ongorora: a: Kana yakashandiswa qPCR/qRT-PCR, iyo fluorescent probe inofanira kuwedzerwa mune reaction system.Kazhinji, yekupedzisira primer concentration ye 0.2 μM inogona kupa zvibereko zvakanaka;kana kuita kwekuita kwakashata, iyo primer concentration inogona kugadziridzwa muhuwandu hwe 0.2-1 μM.Kazhinji, iyo probe concentration inogadziriswa muhuwandu hwe0.1-0.3 μM.Concentration gradient miedzo inogona kuitwa kuti uwane yakanakisa musanganiswa weprimer uye probe.
Notes
1.Iyo yakakwana yekuita tembiricha yeTS-UNG enzyme yakadzikira, uye inogona kugadzirwa mumhando ye20 ℃ ~ 37 ℃, iyo dosage ye enzyme uye nguva yekupindura inogona kugadzirwa muhuwandu hwe0.1 ~ 0.5 U, 5 ~ 10 min;uye enzyme inogona kuvharwa mukuita reverse transcription.
2.Inokodzera PCR uye RT-PCR kudzivirira kusvibiswa.
3.Dzivisa chando chinogara chichinyunguduka, uye usafumure kushanduka kukuru kwetembiricha.
4.Majini akasiyana kuti akwidziridzwe ane akasiyana mashandisiro e dUTP uye senitivity kune UNG enzyme, saka, kana kushandiswa kweUN system kunotungamirira kukudzikira kwekunzwa kwekuona, maitiro ekuita anofanirwa kugadziridzwa uye kugadziridzwa, kana iwe uchida rubatsiro rwehunyanzvi, ndapota taura. kambani yedu.
