Nheyo dzeNucleic Acid Synthesis

Iyo nucleic acid synthesis inoitwa uchishandisa iyo yakasimba phase sialic amide triglyceride nzira, iyo iyo 3 ′ kuguma kweDNA inovharika pane yakasimba chikamu substrate uye nucleotides inowedzerwa mu 3 ′ kusvika 5 ′ kutungamira kusvika yaunoda DNA chidimbu chagadzirwa. .Izvi zvakasiyana kubva kuDNA synthesis nekushandiswa kweDNA polymerase.

Iyo 3'kuguma kwenheyo yekutanga haigadziriswi paCPG panguva yekubatanidza, 5'-OH yenheyo inotevera inodzivirirwa ne di-p-tolyl trityl DMT, boka reamino riri pachigadziko rinodzivirirwa nebenzoic acid uye 3′ -OH inobva yaitwa neamino phosphate compound.1 base ye5 5'-OH ye1 base uye 3'-OH yeinotevera base inoumba phosphite triglyceride, iyo inobva yaiswa oxidine kune phosphate triglyceride, inodzivirira pachigadziko chechipiri 5'-OH inobviswa ne. kuwedzera dichloroacetic acid DM cycles kuburikidza nekuwedzera kwechigadziko chinotevera, uye mushure mekugadzirwa kwekudzivirira pa5'-OH inobviswa neine acid ine simba Chimedu chinobviswa kubva kune yakasimba resin ne concentrated ammonium hydroxide, iyo inodzivirira inobviswa kubva pachigadziko. ine concentrated ammonium hydroxide pasi pekupisa, iyo ammonium hydroxide inobviswa, chidimbu chinoputika chakaomeswa, uye nucleic acid inonatswa ne liquid chromatography kana PEJI.

Matanho eOligo Synthesis

Deblocking

The5 yekupedzisira DMT inodzivirira boka inobviswa nekuwedzera TCA mhinduro.

Activation

Kusanganisa activator neoligonucleotide monomer kuumba inoshanda oligonucleotide monomer yepakati.

Coupling

Iyo 5-terminal hydroxyl boka rinosangana neiyo inoshanda oligonucleotide yepakati mune anti-generative condensation reaction kuti iite isina kugadzikana phosphite triglyceride bond.

Capping 

Kuwedzerwa kwecapping agent inoita acetylation capping reaction neakawandisa hydroxyl mapoka asina kubatikana mune condensation reaction.

Oxidation 

Kuwedzerwa kweiodine mhinduro ine mvura inopindirana nekusagadzikana phosphite zvisungo neoxidation kuumba yakagadzikana phosphoric acid triglyceride bond.

Cyanoethyl Cleavage

Iyo nucleic acid chigadzirwa mushure mekugadzirisa, pamwe nekuwedzera kweDEA mhinduro yekubvisa cyanoethyl boka kubva kune phosphate triglyceride bond.

Yakacheneswa & Kudzivirirwa

Iyo nucleic acid zvimedu zvakatsemurwa kubva kune yakasimba chikamu chinotakura nekuwedzera concentrated ammonium hydroxide solution uyezve kupisa kubvisa mapoka ekudzivirira kubva kumabhondi ehydrogen emabhesi.