Yanayin zafin jiki UNG

Temperatuur Sensitive UNG (TS-UNG) ana samun ta ta hanyar sake haɗawa a cikin E. coli.Enzyme yana haifar da sakin uracil kyauta daga DNA mai dauke da uracil guda-da-biyu kuma baya aiki da RNA.Idan aka kwatanta da na al'ada UNG enzyme na E. coli gene asalin, TS-UNG enzyme yana da mafi girma aiki a low yanayin zafi (20 ℃ ~ 37 ℃) kuma yana da zafin jiki m da sauƙi a kunna (50 ℃), guje wa lalata dUTP-dUTP amplification. samfurori a zafin jiki ta wurin ragowar ayyukan da zai iya kasancewa bayan rashin kunna aikin UNG na al'ada.Sabili da haka, TS-UNG enzyme ba kawai dace da amsawar rigakafin cutar PCR ba, amma kuma yayi daidai da shirin haɓakawa na RT-PCR kuma ana iya amfani dashi a cikin rigakafin kamuwa da cutar RT-PCR.

Aikace-aikacen da aka ba da shawarar

Ƙarfafa Rigakafin Ƙira

Yanayin Ajiya

-20 ° C don ajiya na dogon lokaci, ya kamata a gauraye da kyau kafin amfani, kauce wa daskare-narke akai-akai.

Ma'ajiyar ajiya

20 mM Tris-HCl (pH 7.5), 100 mM NaCl, 0.1 mM EDTA, 1 mM DTT, Stabilizer, 50% Glycerol.

Ma'anar Naúrar

Adadin enzyme da ake buƙata don ƙasƙantar da 1µg na DNA mai ɗaure guda ɗaya mai ɗauke da sansanonin dU a cikin awa 1 a 37°C shine raka'a 1 na aiki (U).

Kula da inganci

1.SDS-PAGE tsarkin electrophoretic fiye da 98%

2.Ayyukan lalata, sarrafa tsari-zuwa-tsari, kwanciyar hankali

3.Babu wani aiki na nuclease, babu exogenous endonuclease ko exonuclease gurbatawa.

Umarni

Lura: a: Idan an yi amfani da shi don qPCR/qRT-PCR, ya kamata a ƙara binciken mai kyalli a cikin tsarin amsawa.Yawancin lokaci, ƙaddamarwa na ƙarshe na 0.2 μM na iya ba da sakamako mai kyau;lokacin da aikin amsawa ya kasance mara kyau, za'a iya daidaita ƙaddamar da ƙaddamarwa a cikin kewayon 0.2-1 μM.Yawancin lokaci, an inganta ƙaddamar da bincike a cikin kewayon 0.1-0.3 μM.Za a iya yin gwaje-gwajen hankali don nemo mafi kyawun haɗe-haɗe na firamare da bincike.

Bayanan kula

1.A mafi kyau duka dauki zazzabi na TS-UNG enzyme ne in mun gwada da low, kuma shi za a iya inganta a cikin kewayon 20 ℃ ~ 37 ℃, da sashi na enzyme da dauki lokaci za a iya inganta a cikin kewayon 0.1 ~ 0.5 U, 5 ~ 10 min;kuma ana iya kunna enzyme a cikin tsarin jujjuya rubutu.

2.Ya dace da PCR da RT-PCR don hana kamuwa da cuta.

3.Guji daskare-narke akai-akai, kuma kar a fallasa ga manyan canjin yanayin zafi.

4.Dabbobi daban-daban da za a haɓaka suna da tasirin amfani daban-daban na dUTP da hankali ga UNG enzyme, sabili da haka, idan amfani da tsarin UNG ya haifar da raguwar ganewar ganewa, ya kamata a daidaita tsarin amsawa da ingantawa, idan kuna buƙatar goyon bayan fasaha, tuntuɓi. kamfaninmu.