Mabda'a aasaasiga ah ee isku dhafka DNA

Isku-dubaridka DNA-da kiimikaadku wuxuu ku salaysan yahay istiraatijiyadda isku-darka wejiga adag iyo kimistariga fosfooraska.Si ka duwan habka DNA-ga noolaha, walxaha ku jira isku-darka DNA-ga kiimikada waa DMT (4, 4-dimethoxytril) iyo phosphoramidite modified deoxyriboxyside, sida ku cad sawirka 1. DNA DNA-da waxaa lagu fuliyaa taageerada adag (waxaan ku siin karnaa noocyo kala duwan oo ah)Tiirka Oligo Synthesisie, CPG ( galaas dalool la kantaroolo) iyo PS (polystyrene)DNA/RNA synthesizer, oo ah qalabkayaga ugu muhiimsan, wuxuu leeyahay noocyo kala duwan sida: HY Single Channel Synthesizer, HY-12, HY-192 iyo iwm, jihada synthesis waa min 3 'ilaa 5', iyo nucleotide ayaa lagu soo bandhigay taageerada adag iyada oo loo marayo hal. wareegga synthesis.Wareegga synthesis caadi ahaan waxa uu ka kooban yahay afar tallaabo, Deprotection, Coupling, Capping and Oxidation (Jaantuska 2).Deprotection-ka ayaa loo dejiyay inuu ka saaro kooxda DMT ee taageerada adag ama kooxda 5' hydroxyl ee nucleoside hore, 3% trichloroacetic acid ee dichloromethane ayaa loo isticmaalaa sidii reagent difaac.Dabadeed fosphoramidite-ka la beddelay ee deoxyriboxyside ayaa loo oggolaaday inay soo bandhigto kooxda hydroxyl iyadoo la kaashanayo activator, ie 5-ethylthiotetrazole ama 4, 5-dicyanoimidazole, samaynta fosfitetrister (III), oo waxay xaqiiqsatay tallaabada Isku-xidhka.Tallaabo Capping ah ayaa la sameeyaa si loo xakameeyo kooxda hydroxyl ee aan la falgelin inta lagu jiro tallaabada isku xirka, si loo yareeyo samaynta taxanaha cilladaha aan loo baahnayn.Ugu dambeyntii, Fosfitetrister-ka aan degganeyn (III) ayaa oksaydhaysan si ay kiimikaad fosfortriester deggan (V) oo leh iodine sida oksaydh ee hadda pyridine.

DNA-da la isku daray ayaa laga yaabaa inay ka soo baxdo taageerada adag ee aminolysis, kooxda 2-cyanoethyl ee la ilaaliyo ee fosfortriester iyo amide ee nucleobase waa la jajabiyaa isku mar, Taarikada Synthesis iyo Synthesis Columns ee racks ayaa si toos ah loogu dhejiyaa qolka falcelinta. ee ahQalabka Ilaalinta.DNA-da cayriin waxaa diyaarin kara HPLC, electrophoresis iyo OPC haddii la rabo, waxaan sidoo kale ku siin karnaaQalabka Sifayntaee Processing ka dib.