Preparation of polydeoxynucleotides linked to a solid support by coupling CNBr-activated cellulose with 5'-NH2-terminated oligo and poly(pdT)'s.
Synthetically-prepared 5'-NH2-dT(pdT)n oligomers (66,n=4 or 7) were immobilized on cyanogen bromide activated cellulose. The influence of temperature, pH, and ionic strength on the rate of the coupling process was studied. The oligomer 5'-NH2-DT(pdT)8 could be elongated enzymatically to the polymers 5'-NH2-dT(pdT)n (n=20, 51 and 84), which could be immobilized on cellulose. The cellulose-NH-dT(pdT)84 polymer thus obtained could be assembled to a new solid-state polymer e.g. poly(dA)290 . poly(/3H/dT)200, poly(dT)85-cellulose which, in turn, was a very convenient substrate for assaying DNA-ligase.
D002482:Cellulose; D055598:Chemical Phenomena; D002621:Chemistry; D003488:Cyanogen Bromide; D007700:Kinetics; D008722:Methods; D008970:Molecular Weight; D009838:Oligodeoxyribonucleotides; D009841:Oligonucleotides; D011071:Poly T; D011089:Polydeoxyribonucleotides; D013696:Temperature; D013938:Thymidine Monophosphate
D009838:Oligodeoxyribonucleotides; D009841:Oligonucleotides; D011089:Polydeoxyribonucleotides; D011071:Poly T; D013938:Thymidine Monophosphate; D002482:Cellulose; D003488:Cyanogen Bromide