Ku-DNA evamile, i-RNA kanye nokuhlanganiswa kwama-nucleic acid okungewona awemvelo, isinyathelo Sokuvikela kanye Nokuhlanganisa sidlala indima ebalulekile.
Isinyathelo Sokuvikela siwukukhipha iqembu le-DMT ekusekelweni okuqinile noma iqembu le-5' hydroxyl ku-nucleoside yangaphambilini ene-organic acid, futhi kudalule iqembu le-hydroxyl ngesinyathelo esilandelayo sokuhlanganisa.I-3% trichloroacetic acid ku-dichloromethane noma i-toluene isetshenziswa kakhulu ukwenza isinyathelo sokuvikela.Ukuhlushwa kwe-trichloroacetic acid kanye nesikhathi sokuvinjelwa (isikhathi sokuvimbela) kubusa ukuhlanzeka kwemikhiqizo yokugcina.Ukugxilisa ingqondo okuphansi nesikhathi esinganele sokuvimba sishiya iqembu le-DMT elingaphenduliwe, elinciphisa isivuno futhi likhulise ukungcola okungafuneki.Isikhathi eside sokuvimba singase siholele i-depurine yokulandelana okuhlanganisiwe, kwakhe ukungcola okungalindelekile.
Isinyathelo Sokuhlanganisa siyazwela kumthamo wamanzi ezincibilikisi kanye nomswakama osemoyeni.Ukuhlangana kwamanzi ekuhlanganiseni kufanele kube ngaphansi kuka-40 ppm, kangcono ngaphansi kuka-25 ppm.Ukugcina isimo se-anhydrous synthesis, ukuhlanganiswa kwe-nucleic acid kufanele kwenziwe endaweni enomswakama ophansi, ngakho-ke sincoma ukuthi amakhasimende ethu asebenziseIzisetshenziswa ezincibilikisiwe zama-Amidites, engancibilikisa iPhosphoramidite eyimpuphu noma enamafutha ku-anhydrous acetonitrile ukugwema ukuthintana nomoya.
Njengoba ukuncibilika kwe-phosphoramidites kungcono esimweni esingewona amanzi, kanye nezicupho zamangqamuzana ukuheha amanzi okulandela umkhondo kuma-reagents nama-amidite, kuyadingeka ukulungisaIzicupho zamangqamuzana.Sincoma u-2 g we-subsieve kumabhodlela e-reagent angu-50-250ml, u-5g kumabhodlela e-reagent angu-250-500ml, u-10g kumabhodlela anyakazayo angu-500-1000ml, kanye no-20g kumabhodlela anyakazayo angu-1000-2000ml.
Ukuncibilika kwe-phosphoramidites kufanele kwenziwe ngaphansi komkhathi ongasebenzi, futhi ukushintshwa kwama-activator reagents kanye ne-acetonitrile kufanele kuqedwe ngesikhathi.I-Capping and Oxidation reagents kufanele isetshenziswe ngokushesha ngangokunokwenzeka, ama-reagents avuliwe anikeza impilo yeshalofu encane, nomsebenzi omncane ngesikhathi sokuhlanganiswa.
Isikhathi sokuthumela: Aug-09-2022