Ana aiwatar da haɗin acid nucleic ta hanyar amfani da tsarin sialic amide triglyceride mai ƙarfi, ta yadda 3′ ƙarshen DNA ya zama mara motsi a kan madaidaicin lokaci mai ƙarfi kuma ana ƙara nucleotides a cikin 3′ zuwa 5′ shugabanci har sai an haɗa gutsure DNA da ake so. .Wannan ya bambanta da haɗin DNA ta aikace-aikacen DNA polymerase.
Ƙarshen 3' na tushe na farko ba shi da motsi a kan CPG yayin haɗuwa, 5'-OH na tushe na gaba yana kiyaye shi tare da di-p-tolyl trityl DMT, ƙungiyar amino a kan tushe an kiyaye shi tare da benzoic acid da 3' -Akan kunna OH tare da hadadden amino phosphate.1 tushe na 5 5'-OH na 1 tushe da 3'-OH na gaba tushe samar da phosphite triglyceride, wanda aka sa'an nan oxidised tare da aidin zuwa phosphate triglyceride, mai kariya a kan tushe na biyu 5'-OH an cire ta ƙara dichloroacetic acid DM hawan keke ta hanyar ƙari na gaba na gaba, kuma bayan haɗawa an cire mai kariya a kan 5'-OH tare da raunin acid mai rauni An cire guntu daga resin mai ƙarfi tare da ammonium hydroxide mai mahimmanci, an cire mai kariya daga tushe. tare da ammonium hydroxide da aka tattara a ƙarƙashin dumama, ana cire ammonium hydroxide, an bushe guntuwar, kuma ana tsabtace nucleic acid ta ruwa chromatography ko PAGE.
Matakan Oligo Synthesis
Kashewa
The5 ƙarshen ƙungiyar kare DMT an cire ta ta ƙara maganin TCA.
Kunnawa
Haɗa mai kunnawa tare da oligonucleotide monomer don samar da matsakaicin oligonucleotide monomer mai aiki.
Haɗin kai
Ƙungiyar hydroxyl ta 5-terminal tana amsawa tare da tsaka-tsakin oligonucleotide mai aiki a cikin wani maganin haɓakaccen ƙwayar cuta don samar da haɗin gwiwar phosphite triglyceride mara kyau.
Yin rubutu
Ƙarin wakili na capping yana aiwatar da amsawar acetylation capping dauki tare da wuce haddi na hydroxyl ƙungiyoyi waɗanda ba su da hannu a cikin motsin motsa jiki.
Oxidation
Ƙarin maganin iodine mai ɗauke da ruwa yana amsawa tare da haɗin phosphite mara kyau ta hanyar oxidation don samar da barga na phosphoric acid triglyceride.
Cyanoethyl Cleavage
Samfurin acid nucleic bayan haɗuwa, tare da ƙari na DEA bayani don cire ƙungiyar cyanoethyl daga haɗin phosphate triglyceride.
Rage&Kariya
An wargaje guntuwar acid nucleic daga ƙaƙƙarfan mai ɗaukar lokaci ta hanyar ƙara daɗaɗɗen ammonium hydroxide bayani sannan kuma mai zafi don cire ƙungiyoyin kariya daga haɗin hydrogen na sansanonin.
Lokacin aikawa: Nuwamba-21-2022