Nā Kumu o ka Nucleic Acid Synthesis

Hoʻokō ʻia ka synthesis nucleic acid me ka hoʻohana ʻana i ke ʻano paʻa sialic amide triglyceride, kahi i hoʻopaʻa ʻia ai ka hopena 3' o ka DNA ma kahi substrate paʻa paʻa a hoʻohui ʻia nā nucleotides ma ka kuhikuhi 3' a hiki i ka 5' a hiki i ka hana ʻana o ka ʻāpana DNA i makemake ʻia. .He ʻokoʻa kēia mai ka synthesis DNA ma o ka hoʻohana ʻana i ka DNA polymerase.

Hoʻopili ʻia ka hopena 3' o ka base mua ma CPG i ka wā synthesis, ua pale ʻia ka 5'-OH o ka base aʻe me ka di-p-tolyl trityl DMT, ʻo ka hui amino ma ke kumu i pale ʻia me ka benzoic acid a me ka 3' A laila ho'āla ʻia ka -OH me kahi pūhui amino phosphate.1 kumu o ka 5 5'-OH o 1 kumu a me 3'-OH o ke kumu aʻe e hana i ka phosphite triglyceride, a laila hoʻoneʻe ʻia me ka iodine i ka phosphate triglyceride, ua wehe ʻia ka mea pale ma ka waihona lua 5'-OH e e hoʻohui i ka dichloroacetic acid DM pōʻai ma o ka hoʻohui ʻana i ke kumu aʻe, a ma hope o ka synthesis ua wehe ʻia ka mea pale ma ka 5'-OH me kahi waika nāwaliwali Ua hemo ka ʻāpana mai ka resin paʻa me ka ammonium hydroxide concentrated, wehe ʻia ka pale mai ke kumu. me ka ammonium hydroxide concentrated ma lalo o ka hoʻomehana, hoʻoneʻe ʻia ka ammonium hydroxide, maloʻo ka ʻāpana, a hoʻomaʻemaʻe ʻia ka nucleic acid e ka chromatography wai a i ʻole PAGE.

Nā ʻanuʻu o Oligo Synthesis

Hoʻopaʻa ʻia

ʻO kaHoʻopau ʻia ka hui pale DMT hope 5 ma ka hoʻohui ʻana i ka hopena TCA.

Hoʻola

Hoʻohui i ka mea hoʻoulu me ka monomer oligonucleotide e hana i ka monomer oligonucleotide ikaika.

Hoʻopili

Hoʻopili ka hui hydroxyl 5-terminal me ka waena oligonucleotide ikaika i loko o kahi hopena condensation anti-generative e hana i kahi paʻa phosphite triglyceride paʻa ʻole.

Kape ana 

ʻO ka hoʻohui ʻana o kahi mea hana capping e hana i kahi hopena acetylation capping me nā hui hydroxyl keu i komo ʻole i ka hopena condensation.

ʻOke ʻana 

ʻO ka hoʻohui ʻana o ka wai iodine i loko o ka wai e hoʻopili ʻia me nā mea paʻa phosphite paʻa ʻole ma o ka hoʻohemo ʻana e hana i nā mea paʻa phosphoric acid triglyceride paʻa.

ʻO Cyanoethyl Cleavage

ʻO ka huahana nucleic acid ma hope o ka synthesis, me ka hoʻohui ʻana i ka solution DEA e wehe i ka hui cyanoethyl mai ka paʻa triglyceride phosphate.

Hoʻokaʻawale a hoʻopaʻa ʻia

Hoʻokaʻawale ʻia nā ʻāpana nucleic acid mai ka mea lawe paʻa paʻa ma o ka hoʻohui ʻana i kahi solution ammonium hydroxide concentrated a laila hoʻomehana ʻia e wehe i nā pūʻulu pale mai nā paʻa hydrogen o nā kumu.