Mu kaphatikizidwe ka DNA, RNA ndi non-natural nucleic acids, gawo la Deprotection and Coupling limakhala ndi ntchito zofunika kwambiri.
Gawo la Deprotection ndikuchotsa gulu la DMT pa chithandizo cholimba kapena gulu la 5 'hydroxyl pa nucleoside yapitayi ndi organic acid, ndikuwulula gulu la hydroxyl pa sitepe yotsatira yolumikizana.3% ya trichloroacetic acid mu dichloromethane kapena toluene imagwiritsidwa ntchito kwambiri poletsa chitetezo.Kuchuluka kwa trichloroacetic acid ndi nthawi yochotsa chitetezo (nthawi yotsekera) zimalamulira chiyero cha zinthu zomaliza.Kutsika kochepa komanso nthawi yosakwanira yotseketsa kumasiya gulu la DMT losakhudzidwa, lomwe limachepetsa zokolola ndikuwonjezera zonyansa zosafunikira.Kutsekeka kwa nthawi yayitali kumatha kutsogoleretsa depurine wazinthu zophatikizika, kupanga zonyansa zosayembekezereka.
Gawo la Coupling limakhudzidwa ndi madzi omwe ali mu zosungunulira komanso chinyezi chomwe chili mumlengalenga.Kuchuluka kwa madzi mu kaphatikizidwe kuyenera kukhala kosakwana 40 ppm, bwino kuchepera 25 ppm.Kusunga chikhalidwe cha anhydrous synthesis, kaphatikizidwe ka nucleic acids kuyenera kuchitidwa pamalo pomwe chinyezi chochepa, chifukwa chake timalimbikitsa makasitomala athu kuti agwiritse ntchitoZida Zosungunuka za Amidites, yomwe imatha kusungunula Phosphoramidite ya ufa kapena mafuta mu anhydrous acetonitrile kuti asagwirizane ndi mpweya.
Popeza kupasuka kwa phosphoramidites ndi bwino pa sanali madzi, ndi misampha maselo adsorb kufufuza madzi mu reagents ndi amidite, m'pofunika kukonzekeraMisampha ya Molecular.Timalimbikitsa 2 g subsieve mabotolo 50-250ml reagent, 5g mabotolo 250-500ml reagent, 10g mabotolo 500-1000ml reagent, ndi 20g 1000-2000ml mabotolo reagent.
Kusungunuka kwa phosphoramidites kuyenera kuchitika pansi pamlengalenga, ndipo m'malo mwa activator reagents ndi acetonitrile kuyenera kutha munthawi yake.Ma reagents a Capping ndi Oxidation ayenera kugwiritsidwa ntchito posachedwa, ma reagents otsegulidwa amapereka moyo wa alumali wocheperako, komanso ntchito yocheperako panthawi ya kaphatikizidwe.
Nthawi yotumiza: Aug-09-2022