To begin, obtain folic acid or choline-deficient feed from commercial rodent food manufacturers. The special diet contains 0.3 milligrams per kilogram of folic acid, or 300 milligrams per kilogram of choline by tartrate. Maintain a female mouse on this special diet for four to six weeks.
Later, when the offspring are weaned from the mother, Place them on a folic acid or choline-deficient diet as well as a control diet. After euthanizing the offspring, collect the blood sample and liver tissue for biochemical analysis. Add the blood to an EDTA-coated tube and centrifuge the tube to isolate the blood plasma for determining the concentration of one-carbon metabolites.
Then, transfer the cut liver tissue samples into a cryo-storage tube for snap freezing. The baseline and weekly weights of female mice on control, choline, or folic acid-deficient diets during the four weeks before mating showed no significant difference. The maternal one-carbon metabolite concentrations were slightly low in the plasma of mice subjected to folic acid-deficient diet as compared to the control mice.