Nucleic acid synthesis e etsoa ho sebelisoa mokhoa o tiileng oa sialic amide triglyceride, moo karolo ea 3' ea DNA e sa sebetseng holim'a karolo e tiileng ea karoloana 'me li-nucleotide li kenngoa ka tsela ea 3' ho ea ho 5' ho fihlela karolo e lakatsehang ea DNA e kopantsoe. .Sena se fapane le tlhahiso ea DNA ka tšebeliso ea DNA polymerase.
Qetello ea 3 ea setsi sa pele ha e sebetse ho CPG nakong ea ho qaptjoa, 5'-OH ea setsi se latelang se sirelelitsoe ka di-p-tolyl trityl DMT, sehlopha sa amino se motheong se sirelelitsoe ka benzoic acid le 3′. -OH e ntan'o sebelisoa ka motsoako oa amino phosphate.Motheo o le mong oa 5 5'-OH oa motheo oa 1 le 3'-OH oa motheo o latelang o etsa phosphite triglyceride, eo ka nako eo e nang le oxidized ka iodine ho phosphate triglyceride, mosireletsi motheong oa bobeli 5'-OH o tlosoa ke ho eketsa dichloroacetic acid DM cycles ka ho eketsoa ha setsi se latelang, 'me ka mor'a ho kopanya mosireletsi ho 5'-OH o tlosoa ka acid e fokolang Sekhechana se khaotsoe ho tloha ho resin e tiileng ka concentrated ammonium hydroxide, mosireletsi o tlosoa motheong. ka concentrated ammonium hydroxide tlas'a ho futhumala, ammonium hydroxide e tlosoa, sekhechana se omisitsoe, 'me nucleic acid e hloekisoa ke liquid chromatography kapa PAGE.
Mehato ea Oligo Synthesis
Deblocking
TheSehlopha sa 5 se sireletsang DMT se tlosoa ka ho eketsa tharollo ea TCA.
Tshebetso
Ho kopanya activator le oligonucleotide monomer ho theha oligonucleotide monomer e sebetsang lipakeng.
Ho kopanya
Sehlopha sa 5-terminal hydroxyl se sebetsana le oligonucleotide e sebetsang ka hare ho anti-generative condensation reaction ho theha bond e sa tsitsang ea phosphite triglyceride.
Capping
Ho eketsoa ha capping agent ho etsa acetylation capping reaction ka lihlopha tse feteletseng tsa hydroxyl tse sa ameheng karabelong ea condensation.
Oxidation
Ho eketsoa ha tharollo ea iodine e nang le metsi e sebetsana le litlamo tse sa tsitsang tsa phosphite ka oxidation ho theha maqhama a tsitsitseng a phosphoric acid triglyceride.
Cyanoethyl Cleavage
Sehlahisoa sa nucleic acid ka mor'a ho kopanya, ka ho eketsoa ha tharollo ea DEA ho tlosa sehlopha sa cyanoethyl ho tloha ho phosphate triglyceride bond.
Khiloe & Tšireletso
Likaroloana tsa asiti ea nucleic li petsoha ho sejari sa mohato o tiileng ka ho eketsa tharollo ea ammonium hydroxide e tebileng ebe e futhumatsoa ho tlosa lihlopha tse sirelletsang litlamong tsa hydrogen tsa metheo.
Nako ea poso: Nov-21-2022