Isku-darka aashitada nucleic-ka waxaa lagu fuliyaa iyadoo la adeegsanayo habka adag ee sialic amide triglyceride, halkaas oo 3 ′ dhamaadka DNA-da aan dhaqdhaqaaq lahayn on substrate weji adag iyo nucleotides ayaa lagu daraa jihada 3′ ilaa 5′ ilaa inta la rabo in la isku daro DNA-da la rabo. .Tani way ka duwan tahay isku-darka DNA-ga iyadoo la adeegsanayo polymerase DNA.
Dhammaadka 3′ ee saldhigga ugu horreeya waxa lagu ilaaliyaa CPG inta lagu jiro isku-dhafka, 5′-OH ee saldhigga xiga waxaa lagu ilaaliyaa di-p-tolyl trityl DMT, kooxda amino ee saldhigga waxaa lagu ilaaliyaa benzoic acid iyo 3′ -OH ka dib waxaa lagu hawlgeliyaa iskudarka fosfaafiska amino.1 saldhig ee 5 5'-OH ee 1 saldhiga iyo 3'-OH ee saldhiga xiga waxay sameeyaan fosfit triglyceride, ka dibna oksaydhka leh iodine ilaa phosphate triglyceride, ilaaliyaha saldhiga labaad 5'-OH waxaa laga saarayaa by ku darida dichloroacetic acid wareegyada DM iyada oo lagu daro saldhigga xiga, ka dib marka la isku daro ilaaliyaha 5'-OH waxaa laga saarayaa asiidh daciif ah Jajabka ayaa ka go'aya xajinta adag ee leh ammonium hydroxide oo xoogga leh, difaaca ayaa laga saaraa saldhigga. oo leh ammonium hydroxide oo kuleyl ah, ammonium hydroxide waa laga saarayaa, jajabkii waa la qalajiyey vacuum, iyo nucleic acid waxaa lagu nadiifiyaa chromatography dareere ah ama PAGE.
Tallaabooyinka Oligo Synthesis
Joojinta
The5 dhamaadka DMT kooxda ilaalinta waa la saaray iyadoo lagu daray TCA xal.
Firfircoonida
Isku darka firfircoonida iyo monomer oligonucleotide si loo sameeyo dhexdhexaadiyaha oligonucleotide monomer firfircoon.
Isku-xidhka
Kooxda 5-terminal hydroxyl waxay la falgalaan dhexdhexaadiyaha oligonucleotide firfircoon ee falcelinta ka-hortagga uumiga si ay u sameeyaan curaarta phosphite triglyceride aan degganayn.
duubista
Ku darida wakiilka daboolka waxay qabataa falcelinta daboolka acetylation-ka oo ay la socdaan kooxaha hydroxyl ee xad-dhaafka ah ee aan ku lug lahayn falcelinta uumiga.
Oxidation
Ku darida xalalka iodine ee ka kooban biyaha waxay la falgashaa curaarta fosfit-ka ee aan xasilloonayn iyadoo ay ogsaydheyntu u samaynayso curaarta phosphoric acid triglyceride deggan.
Cyanoethyl Cleavage
Badeecada aashitada nucleic ka dib markii la isku daro, iyadoo lagu daro xalka DEA si looga saaro kooxda cyanoethyl ee phosphate triglyceride bond.
Go'ay&Difaacid
Qaybaha aashitada nukliyeerka ayaa laga soo jeexay side wejiga adag iyada oo lagu darayo xal ammonium hydroxide oo xoog leh ka dibna la kululeeyo si looga saaro kooxaha ilaalinta ee curaarta hydrogen ee saldhigyada.
Waqtiga boostada: Nov-21-2022