One hundred sporadic colorectal cancer patients, 100 newborns and 100 adults without a history of malignant disease, were investigated to determine the allele frequencies and genotype distribution of the SULT1A1 Arg213His variant in the population of the Republic of Macedonia (Table 1).
      Genomic DNA was extracted from whole blood, paraffin embedded tissues and/or fresh tumors, using a QIA­GEN DNA extraction kit and the procedure recommended by the manufacturer (QIAGEN AS, Oslo, Norway). The presence of the variant SULT1A1 gene was determined with a polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) procedure as follows: a fragment of 282 bp including exon 7 of the SULT1A1 gene was amplified by PCR using primers 5’-GGT TGA GGA GTT GGC TCT GC-3’ and 5’-ATG AAC TCC TGG GGG ACG GT-3’ on a GeneAmp PCR System 2700 (Applied BioSystems, Foster City, CA, USA). The PCR conditions included initial denaturation at 95°C for 10 min., 33 cycles of denaturation (95°C for 30 seconds), annealing (62°C for 30 seconds) and extension (72°C for 1 min.), and a 10 min. final extension at 72°C. The PCR products were incubated with HaeII at 37°C overnight. The resulting fragments were resolved by electrophoresis on 2% agarose gels in TBE buffer (0.89 M Ttris-HCl, 0.89 M Boric acid and 0.02 M EDTA-Na2-salt), visualized by ethidium bromide staining and recorded on digital camera. Representative results showing the detection of various genotypes in four samples are shown in Figure 1.
      Comparison of allele frequencies and genotype distributions between patients and controls was done by Chi-square, Fisher’s exact test and Multivariate statistical analysis with Bonferoni correction. The genotype frequencies in patients and controls were tested if they follow the Hardy-Weinberg equilibrium.

Table 1. Some characteristics of the patient and control groups in this study.