Ụkpụrụ nke Nucleic Acid Synthesis

A na-eme njikọ njikọ nucleic acid site na iji usoro siri ike sialic amide triglyceride, nke a na-eme ka njedebe 3 nke DNA ghara imegharị ya na mkpụrụ osisi siri ike na nucleotides na-agbakwunye na 3' ruo 5' direction ruo mgbe a na-ejikọta iberibe DNA achọrọ. .Nke a dị iche na njikọ DNA site na ntinye nke DNA polymerase.

A na-eme ka njedebe nke 3 nke mbụ dị na CPG n'oge njikọ, 5'-OH nke isi na-esote na-echebe ya na di-p-tolyl trityl DMT, amino group na isi na-echebe ya na benzoic acid na 3′ -OH na-arụ ọrụ na amino phosphate compound.1 isi nke 5 5 ′-OH nke 1 isi na 3′-OH nke na-esote isi na-etolite a phosphite triglyceride, nke na-ahụ oxidised na ayodiin ka a phosphate triglyceride, onye na-echebe na nke abụọ isi 5′-OH wepụrụ site na. na-agbakwunye dichloroacetic acid DM cycles site na mgbakwunye nke isi na-esote, na mgbe njikọta ihe nchebe na 5′-OH na-ewepụ ya na acid na-adịghị ike A na-ewepụ iberibe ahụ site na resin siri ike na ammonium hydroxide na-etinye uche, a na-ewepụ ihe nchebe na isi. na ammonium hydroxide na-etinye uche n'okpuru kpo oku, a na-ewepụ ammonium hydroxide, a na-ekpochapụ iberibe ahụ, a na-ehichapụkwa nucleic acid site na chromatography mmiri ma ọ bụ PAGE.

 

Nzọụkwụ Oligo Synthesis

Mgbochi

NkeA na-ewepụ 5 end DMT otu nchekwa site na ịgbakwunye ngwọta TCA.

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Nkwalite

Ịgwakọta onye na-eme ihe na oligonucleotide monomer ka ọ bụrụ etiti oligonucleotide monomer na-arụ ọrụ.

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Njikọta

Otu 5-terminal hydroxyl na-emeghachi omume na etiti oligonucleotide na-arụ ọrụ na mmeghachi omume condensation nke na-eme ka ọ bụrụ njikọ phosphite triglyceride na-adịghị akwụsi ike.

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Nkwudo okwu 

Mgbakwunye nke onye na-ahụ maka capping na-eme mmeghachi omume capping acetylation na oke hydroxyl dị iche iche na-adịghị etinye aka na mmeghachi omume condensation.

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Oxidation 

Mgbakwunye nke ngwọta ayodiin nwere mmiri na-emeghachi omume na njikọ phosphite na-adịghị akwụsi ike site na oxidation iji mepụta njikọ phosphoric acid triglyceride kwụsiri ike.

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Cyanoethyl Cleavage

Ngwaahịa nucleic acid mgbe njikọta, yana mgbakwunye nke ngwọta DEA iji wepụ otu cyanoethyl na njikọ phosphate triglyceride.

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Ekewara&nchekwa

A na-agbaji iberibe acid nucleic site na onye na-ebu akụkụ siri ike site na ịgbakwunye ngwọta ammonium hydroxide na-ekpo ọkụ wee kpoo ọkụ iji wepụ otu ndị na-echebe site na njikọ hydrogen nke bases.

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Oge nzipu: Nov-21-2022