Enzymatic synthesis of selectively deuterated poly [d(A-T)].poly[d(A-T)]

Ingrid Parrot, Valerie Laux, and Trevor Forsyth

Keele University and Institut Laue Langevin

Polymerisation reaction (analytical scale)

8 ml hydrogenated dATP 50mM

8 ml deuterated dTTP 50 mM

16 ml Tris 600 mM pH 8

124 ml sterile H2O

12 ml MgCl2 100 mM

26 ml inorganic pyrophosphatase 0.0015 u/ml

4 ml primer poly[d(A-T)].poly[d(A-T] @ 1mg/ml

2 ml DNA Pol I Klenow 5 u/ml

 

Incubate at 37o C

 
Agarose (1%) gel electrophoresis  showing the time course of the polymerisation reaction and the size of the final product. Size exclusion chromatography illustrating the high efficiency of the polymerisation reaction

For further information, contact Ingrid Parrot (parrot@ill.fr)