High‐resolution single‐stranded DNA analysis on 4.5 cm plastic electrophoretic microchannels

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Abstract
Plastic microchannels (4.5 cm long) fabricated from an etched glass master were tested for high‐resolution single‐stranded DNA analysis. Using replaceable denaturing linear polyacrylamide as sieving matrix, one‐color separation of a fragment sizing standard with single‐base resolution (R > 0.5) was achieved up to 275 bases. Two‐color sizing analysis of four loci short tandem repeat (STR) allelic ladder (CSF1PO, TPOX, TH01, vWA) with single‐base resolution (R = 0.62) on TH01 alleles 9.3 (198 bp) and 10 (199 bp) was demonstrated. An average standard deviation of ± 0.06 bp and ±0.11 bp in sizing 32 alleles of the CTTv ladder was attained between runs and between channels, respectively. Four‐color sequencing separation of a terminator sequencing standard showed a base‐calling accuracy of 99.1% out to 320 bases in 13 min.