Parallel analysis of amplicons at multiple temperatures. (a) Using a manifold (see Fig. 1a for instrumental details) the sample was split and directed to four monolithic capillary columns kept at four different temperatures. (b) The same samples had been analyzed previously serially on a commercially available micropellicular column support at 52°C and 57°C. Experimental conditions: (a) columns, 4 × monolithic PS/DVB, 60 × 0.2 mm i.d.; mobile phase, (A) 100 mM TEAA at pH 7.0, (B) 100 mM TEAA at pH 7.0, 25% acetonitrile; linear gradient, 45%–61% B in 8.0 min; flow rate, 2.0–3.0 μl/min; temperature, 51°C, 53°C, 55°C, and 57°C, respectively; detection, LIF, emission monitored at 525 nm; injection volume, 1 μL each; (b) column, DNASep, 50 × 4.6 mm i.d.; mobile phase, (A) 100 mM TEAA at pH 7.0, (B) 100 mM TEAA at pH 7.0, 25% acetonitrile; linear gradient, 58%–64% B in 3.0 min at 52°C, and 54%–60% B in 3.0 min at 57°C; flow rate, 900 μL/min; detection, UV, 254 nm; injection volume: 8 μL each. Samples: one homozygous and three different heterozygous samples comprising exon 39 and flanking noncoding regions of ATM.