Nheyo yekutanga yeDNA synthesis

Iyo kemikari DNA synthesis yakavakirwa pane yakasimba-chikamu synthesis zano uye phosphoramidite chemistry.Yakasiyana neiyo biological DNA synthesis, zvinhu zviri mumakemikari DNA synthesis iDMT (4, 4-dimethoxytrityl) uye phosphoramidite yakagadziridzwa deoxyriboxyside, sezvakaratidzwa mumufananidzo 1. Iyo DNA synthesis inoitwa pane yakasimba rutsigiro (tinogona kupa akasiyanaOligo Synthesis Column), kureva CPG (inodzorwa pore girazi) uye PS (polystyrene), uye iyo yese synthesis inoitwa paneDNA/RNA synthesizer, inova midziyo yedu huru, ine modhi yakasiyana yakadai se: HY Single Channel Synthesizer, HY-12, HY-192 uye nezvimwe, iyo synthesis nzira inobva pa3' kusvika 5', uye nucleotide inounzwa kune yakasimba rutsigiro kuburikidza neimwe. synthesis cycle.A anowanzo synthesis cycle ine matanho mana, Deprotection, Coupling, Capping uye Oxidation (Mufananidzo 2).Iyo Deprotection yakagadzirirwa kubvisa boka reDMT parutsigiro rwakasimba kana 5' hydroxyl boka pane yapfuura nucleoside, iyo 3% trichloroacetic asidhi mu dichloromethane inoshandiswa seyekudzivirira reagent.Ipapo phosphoramidite yakagadziridzwa deoxyriboxyside yakabvumidzwa kuratidza hydroxyl boka nerubatsiro rwe activator, kureva 5-ethylthiotetrazole kana 4, 5-dicyanoimidazole, kugadzira phosphitetriester (III), uye akaziva Coupling danho.Danho reCapping rinoitwa kuvharisa iro risingaite hydroxyl boka panguva yekubatanidza nhanho, kudzikisa kuumbwa kwezvisina kudiwa chirema.Pakupedzisira, iyo isina kugadzikana phosphitetriester (III) yakasvibiswa kumakemikari yakagadzikana phosphortriester (V) ine iodine seyo oxidant mune iripo yepyridine.

Iyo yakagadzirwa DNA inogona kuve yakatsemuka kubva pakutsigirwa kwakasimba neaminolysis, iyo 2-cyanoethyl boka rakadzivirirwa pane phosphortriester uye iyo amide pane nucleobase inotsemurwa panguva imwe chete, iyo Synthesis Plates uye Synthesis Columns mumaracks anoiswa zvakananga mukamuri yekupindura. yeDeprotection Equipment.Crude DNA inogona kugadzirwa neHPLC, electrophoresis uye OPC pazvinenge zvichidiwa, tinogona kukupawoZvishandiso zvekuchenesayePost-processing.