Kwi-DNA eqhelekileyo, i-RNA kunye ne-non-natural nucleic acids' synthesis, inyathelo lokuKhusela kunye nokuDibana lidlala indima ebalulekileyo.
Isinyathelo sokuKhuselwa kukususa iqela le-DMT kwinkxaso eqinile okanye i-5 'iqela le-hydroxyl kwi-nucleoside yangaphambili kunye ne-organic acid, kwaye iveze iqela le-hydroxyl kwisinyathelo esilandelayo sokudibanisa.I-3% ye-trichloroacetic acid kwi-dichloromethane okanye i-toluene isetyenziswa kakhulu ukwenza inyathelo lokukhusela.Ukuxinwa kwe-trichloroacetic acid kunye nexesha lokukhusela (ixesha lokutshatyalaliswa) lilawula ukucoceka kweemveliso zokugqibela.Ukugxininiswa okuphantsi kunye nexesha elaneleyo lokunciphisa lishiya iqela le-DMT elingaphendulwanga, elinciphisa isivuno kunye nokwandisa ukungcola okungafunekiyo.Ixesha elide lokuvala lingakhokelela kwi-depurine yolandelelwano oluhlanganisiweyo, yenze ubumdaka obungalindelekanga.
Isinyathelo sokuDityaniswa sinovakalelo kumxholo wamanzi wezinyibilikisi kunye nokufuma emoyeni.Ukuxinwa kwamanzi kwi-synthesis kufuneka kube ngaphantsi kwe-40 ppm, ngcono ngaphantsi kwe-25 ppm.Ukugcina imeko ye-anhydrous synthesis, i-nucleic acids 'synthesis kufuneka yenziwe kwindawo yokufuma okuphantsi, ke sicebisa umthengi wethu ukuba asebenziseIzixhobo ezinyibilikileyo zeAmidites, enokuthi ichithe i-Phosphoramidite engumgubo okanye i-oily kwi-acetonitrile engenamanzi ukuphepha ukudibanisa nomoya.
Ekubeni ukunyibilika kwe-phosphoramidites kungcono kwimeko engeyoyamanzi, kunye nemigibe yeemolekyuli ukubhengeza umkhondo wamanzi kwi-reagents kunye ne-amidite, kuyimfuneko ukulungiselelaImigibe yeemolekyuli.Sincoma i-2 g ye-subsieve kwiibhotile ze-reagent ye-50-250ml, i-5g kwiibhotile ze-reagent ye-250-500ml, i-10g ye-500-1000ml iibhotile ze-reagent, kunye ne-20g ye-1000-2000ml iibhotile ze-reagent.
Ukuchithwa kwe-phosphoramidites kufuneka kuqhutywe phantsi kwe-atmosphere ye-inert, kwaye ukutshintshwa kwe-activator reagents kunye ne-acetonitrile kufuneka kugqitywe ngexesha.I-Capping kunye ne-Oxidation reagents kufuneka isetyenziswe ngokukhawuleza, ii-reagents ezivuliwe zinika ubomi obuncinci beshelufu, kunye nomsebenzi omncinci ngexesha lokudibanisa.
Ixesha lokuposa: Aug-09-2022