Ko te kaupapa matua o te whakahiato DNA

Ko te whakahiato DNA matū i ahu mai i te rautaki whakahiato-waahanga totoka me te matū phosphoramidite.He rereke mai i te whakahiato DNA koiora, ko nga mea i roto i te whakahiato DNA matū ko DMT (4, 4-dimethoxytrityl) me te deoxyriboxyside kua whakarereketia te phosphoramidite, e whakaatuhia ana i te Whakaahua 1. Ka mahia te whakahiato DNA i runga i te tautoko totoka(ka taea e matou te tuku momo momoOligo Synthesis Column), arā, ko te CPG (karaihe pore whakahaere) me te PS (polystyrene), ka mahia te whakahiatotanga katoa i runga i teKo te DNA/RNA synthesizer, Ko a maatau taputapu matua, he tauira rereke penei: HY Single Channel Synthesizer, HY-12, HY-192 me etahi atu, ko te ahunga whakahiato mai i te 3' ki te 5', ka whakauruhia he nucleotide ki te tautoko totoka ma te kotahi huringa whakahiato.Kei roto i te huringa whakahiatotanga e wha nga kaupae, Te Tiakitanga, Te Hononga, Te Whakakapi me te Haahi (Whakaahua 2).Kua whakaritea te Deprotection ki te tango i te roopu DMT i runga i te tautoko totoka, i te roopu 5' hydroxyl ranei i runga i te nucleoside o mua, ko te 3% waikawa trichloroacetic i roto i te dichloromethane ka whakamahia hei reagent deprotection.Na ka whakaaetia te phosphoramidite deoxyriboxyside whakarerekētia ki te whakaatu rōpū hydroxyl ki te āwhina o te activator, arā 5-ethylthiotetrazole ranei 4, 5-dicyanoimidazole, hanga i te phosphitetriester (III), a ka mohio i te taahiraa Hononga.Ka mahia he mahi Whakakapi hei aukati i te roopu hydroxyl kaore i tauhohehia i te wa e hono ana, hei whakaiti i te hanga o nga raupapa kohakore.Ka mutu, ka whakapourihia te phosphitetriester (III) korekore ki te phosphotriester pumau matū (V) me te iodine hei hāora i roto i te hakari o te pyridine.

Ko te DNA synthesized ka taea te wehe mai i te tautoko totoka na te aminolysis, ko te roopu tiaki 2-cyanoethyl i runga i te phosphotriester me te amide i runga i te nucleobase ka wehea i te wa ano, ko nga Pereti Synthesis me nga Kohanga Whakakotahitanga ka tuu tika ki roto i te ruma tauhohenga. o ngaTaputapu Tangohanga.Ka taea e te HPLC, te electrophoresis me te OPC te whakarite DNA maru, ka taea hoki e matou te whakarato ki a koeTaputapu purengamo te Tukatuka-muri.