°C for 3 s and 60 °C for 30 s. Each reaction was carried out in a total volume of 20 μl containing 1 × Taqman Fast Universal PCR Master mix, 900 nM of each primer and 250 nM probe. A total of 2.5 μl of DNA was added to each reaction. Reactions were carried out in triplicate. Quantitative PCR analysis software supplied by the manufacturer (Applied Biosystems) was used to calculate standard curves and quantify samples. A control probe set within the albumin gene was used as a reference to determine copy number. A ratio of 1 for assay/albumin was viewed as normal copy number.
