Ihame shingiro rya synthesis ya ADN

Imiti ya ADN ya chimique ishingiye ku ngamba zikomeye zo guhuza hamwe na chimie ya fosifora.Bitandukanye na synthesis ya ADN yibinyabuzima, ibikoresho biri muri synthesis ya ADN ya chimique ni DMT (4, 4-dimethoxytrityl) na fosiforamide yahinduwe deoxyriboxyside, nkuko bigaragara ku gishushanyo cya 1. Synthesis ya ADN ikorwa ku nkunga ikomeye (dushobora gutanga ibintu bitandukanyeInkingi ya Oligo), ni ukuvuga CPG (ikirahuri cya pore igenzurwa) na PS (polystirene), kandi synthesis yose ikorerwa kuri aADN / RNA synthesizer, nicyo gikoresho cyacu nyamukuru, gifite icyitegererezo gitandukanye nka: HY Umuyoboro umwe wa Synthesizer, HY-12, HY-192 nibindi, icyerekezo cya synthesis kiva kuri 3 'kugeza 5', kandi nucleotide itangirwa inkunga ikomeye ikoresheje imwe synthesis cycle.Mubisanzwe synthesis cycle ikubiyemo intambwe enye, Deprotection, Coupling, Capping na Oxidation (Ishusho 2).Deprotection yashyizweho kugirango ikureho itsinda rya DMT ku nkunga ikomeye cyangwa itsinda rya 5 'hydroxyl kuri nucleoside yabanjirije iyi, aside 3% ya trichloroacetic muri dichloromethane ikoreshwa nka reagente reagent.Hanyuma fosiforamide yahinduwe deoxyriboxyside yemerewe kwerekana itsinda rya hydroxyl abifashijwemo na activateur, ni ukuvuga 5-Ethylthiotetrazole cyangwa 4, 5-dicyanoimidazole, bagize fosifeti (III), maze bamenya intambwe yo guhuza.Intambwe ya Capping irakorwa kugirango uhagarike hydroxyl groupe idakozwe mugihe cyo guhuza intambwe, kugirango hagabanuke imiterere yinenge idakenewe.Hanyuma, fosifeti itajegajega (III) ihindurwamo okiside ya fosifori ihamye (V) hamwe na iyode nka okiside mugihe cya pyridine.

ADN ikomatanyirijwe irashobora gukurwa mubufasha bukomeye na aminolysis, itsinda rya 2-cyanoethyl ririnzwe kuri fosifori na amide kuri nucleobase yacitse icyarimwe, Isahani ya Synthesis hamwe na Synthesis Inkingi mubitereko bishyirwa mubyumba byabigenewe. Bya iIbikoresho bya Deprotection.ADN yuzuye irashobora gutegurwa na HPLC, electrophoreis na OPC kubisabwa, turashobora no kuguhaIbikoresho byo kwezaKuri Nyuma yo gutunganya.