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Nucleic Acids Research, 1979, Vol. 6, No. 1 247-258
© 1979

Articles

Preparation of polydeoxynucleotides linked to a solid support by coupling CNBr-activated cellulose with 5'-NH2-terminated oligo and poly(pdT)'s*

L. Clerici{dagger}, F. Campagnari{dagger}, J.F.M.de Rooij+ and J.H. van Boom+

{dagger}Laboratory of Biochemistry, Biology Group Ispra D.G. XII, C.E.C. Joint Research Centre, 21020 Ispra (Va), Italy +Gorlaeus Laboratory 2300 RA Leiden, Netherlands

Received October 24, 1978.

Synthetically-prepared 5'-NH2-dT(pdT)noligomers (6b,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)8could be elongated enzymatically to the polymers 5'-NH 2.-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)290poly|3H|dT)200, poly(dT)85-cellulose which, in turn, was a very convenient substrate for assaying DNA-ligase.

*This publication is contribution No. 1556 of the Programme; "Biology, Radiation Protection and Medical Research", Directorate General XII for Research, Science and Education of the Commission of the European Communities, C.E.C.


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