Asalin ka'idar haɗin DNA

Haɗin DNA na sinadarai ya dogara ne akan dabarun haɗin lokaci mai ƙarfi da kuma phosphoramite chemistry.Daban-daban daga halittar DNA kira na halitta, abu a cikin sinadarai DNA kira ne DMT (4, 4-dimethoxytril) da phosphoramidite modified deoxyriboxyside, kamar yadda aka nuna a cikin Hoto 1. A DNA kira ne da za'ayi a kan m goyon baya (za mu iya bayar da daban-daban naginshiƙi na Oligo Synthesiswatau CPG (gilashin pore mai sarrafawa) da PS (polystyrene), kuma ana yin duka kirar akanDNA/RNA synthesizer, wanda shine babban kayan aikin mu, yana da nau'i daban-daban kamar: HY Single Channel Synthesizer, HY-12, HY-192 da dai sauransu, jagorancin kira yana daga 3 'zuwa 5', kuma an gabatar da nucleotide zuwa ingantaccen tallafi ta hanyar daya. sake zagayowar kira.Tsarin sake haɗawa ya ƙunshi matakai huɗu, Deprotection, Coupling, Capping and Oxidation (Hoto na 2).An saita Deprotection don cire ƙungiyar DMT akan ingantaccen tallafi ko ƙungiyar 5' hydroxyl akan nucleoside da ta gabata, ana amfani da 3% trichloroacetic acid a cikin dichloromethane azaman reagent deprotection.Sa'an nan phosphoramidite modified deoxyriboxyside aka yarda da fallasa hydroxyl kungiyar tare da taimakon activator, watau 5-ethylthiotetrazole ko 4, 5-dicyanoimidazole, forming phosphitetriester (III), da kuma gane Coupling mataki.Ana aiwatar da matakin capping don toshe ƙungiyar hydroxyl da ba a yi ba yayin matakin haɗin gwiwa, don rage samuwar jerin lahani da ba a so.A ƙarshe, phosphitetriester mara ƙarfi (III) yana oxidized zuwa sinadarai barga phosphortriester (V) tare da aidin a matsayin oxidant a halin yanzu na pyridine.

DNA da aka haɗa na iya zama cleavage daga ingantaccen goyon baya ta hanyar aminolysis, ƙungiyar kariya ta 2-cyanoethyl akan phosphortriester kuma amide akan nucleobase yana ɓarke ​​​​a lokaci guda, faranti na Ƙarfafawa da Rukunin Rubutun a cikin racks ana sanya su kai tsaye a cikin ɗakin amsawa. naKayayyakin Kariya.Ana iya shirya ɗanyen DNA ta HPLC, electrophoresis da OPC akan buƙata, kuma zamu iya samar muku daKayan aikin tsarkakewadon Post-processing.