Izimiso ze-Nucleic Acid Synthesis

I-nucleic acid synthesis yenziwa kusetshenziswa indlela eqinile yesigaba se-sialic amide triglyceride, lapho isiphetho se-3′ se-DNA singenakunyakaziswa ku-substrate yesigaba esiqinile futhi ama-nucleotide engezwa ohlangothini lwe-3' kuya ku-5' kuze kube yilapho kuhlanganiswa ucezu lwe-DNA olufunekayo. .Lokhu kwehluka ekuhlanganiseni kwe-DNA ngokusetshenziswa kwe-DNA polymerase.

Ukuphela kwe-3' kwesisekelo sokuqala kuvinjelwe ku-CPG ngesikhathi sokuhlanganiswa, i-5'-OH yesisekelo esilandelayo ivikelwe nge-di-p-tolyl trityl DMT, iqembu le-amino esisekelweni livikelwe nge-benzoic acid kanye ne-3′ -OH bese icushwa ngenhlanganisela ye-amino phosphate.Isisekelo esingu-1 sika-5 5'-OH wesisekelo esingu-1 kanye no-3'-OH wesisekelo esilandelayo sakha i-phosphite triglyceride, ebuye ifakwe i-iodine ibe i-phosphate triglyceride, isivikelo esisekelweni sesibili esingu-5'-OH sisuswa ngu. ukwengeza imijikelezo ye-dichloroacetic acid DM ngokungezwa kwesisekelo esilandelayo, futhi ngemva kokuhlanganiswa isivikelo ku-5'-OH sisuswa nge-asidi ebuthakathaka Ingxenye inqanyulwa ku-resin eqinile nge-concentrated ammonium hydroxide, isivikelo siyasuswa esisekelweni. nge-ammonium hydroxide egxilile ngaphansi kokufudumeza, i-ammonium hydroxide iyasuswa, ucezu luyasuswa, futhi i-nucleic acid ihlanzwa nge-chromatography ewuketshezi noma i-PAGE.

Izinyathelo ze-Oligo Synthesis

Iyasusa

IIqembu eli-5 lokuvikela i-DMT lisuswa ngokungeza isisombululo se-TCA.

Ukwenza kusebenze

Ukuxuba i-activator ne-oligonucleotide monomer ukwakha i-oligonucleotide monomer esebenzayo emaphakathi.

Ukuhlanganisa

Iqembu le-5-terminal hydroxyl lisabela ne-oligonucleotide esebenzayo emaphakathi ku-anti-generative condensation reaction ukwakha ibhondi ye-phosphite triglyceride engazinzile.

I-Capping 

Ukwengezwa kwe-ejenti ye-capping kwenza ukusabela kwe-acetylation capping namaqembu e-hydroxyl eyeqile angabandakanyeki ekuphenduleni kwe-condensation.

I-oxidation 

Ukwengezwa kwesisombululo se-iodine esiqukethe amanzi kuhlangana namabhondi e-phosphite angazinzile nge-oxidation ukuze kwakheke amabhondi ezinzile we-phosphoric acid triglyceride.

I-Cyanoethyl Cleavage

Umkhiqizo we-nucleic acid ngemva kokuhlanganiswa, ngokungezwa kwesisombululo se-DEA ukususa iqembu le-cyanoethyl ku-phosphate triglyceride bond.

Kususiwe&Kuvikelwe

Izingcezwana ze-nucleic acid ziklebhuka kusithwali sesigaba esiqinile ngokwengeza isixazululo esigxilile se-ammonium hydroxide bese siyashisisa ukuze kususwe amaqembu avikelayo kumabhondi e-hydrogen ezisekelweni.