To locate the genes that make a substantial contribution to variation in natural dizygotic twinning in humans, large-scale studies are needed. New studies should not stop at DNA genotyping, but collect material that allow gene-expression analysis, transcriptomics, proteomics and endocrinology. In this article we describe a pilot study to examine the feasibility, effectiveness and logistics of large-scale nationwide sample collection in Dutch families in which two or more sisters have given birth to spontaneous dizygotic twins. Pedigree data and addresses from family members of proband mothers were collected by telephone. Blood and urine samples were collected during a home visit, and handled in the afternoon. All participants were bled between 7 a.m. and 10 a.m. after overnight fasting. Blood samples of fertile women with a natural cycle were collected on the second, third or fourth day of their menstrual cycle. The effects of transportation and storage on blood quality, lipids, RNA with and without challenge, lymphocytes and other parameters were examined. Genomic DNA was isolated from blood and cells were immortalized using Epstein–Barr virus. In 78.6% of the women with a natural cycle blood samples were collected on the second, third or fourth day of the menstrual cycle. This percentage is likely to increase with the more dense geographical distribution of participants in the larger population. We conclude that the pilot study demonstrated the feasibility of this protocol to collect good quality of plasma, DNA, RNA and lymphocyte samples by home visits.