High-resolution single-stranded DNA analysis on 4.5 cm plastic electrophoretic microchannels.
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.
Shi
Yining
Y
ACLARA BioSciences, Inc., Mountain View, CA 94043, USA. yshi@ACLARA.com
Anderson
Rolfe C
RC
D000180:Acrylic Resins; D000483:Alleles; D003116:Color; D004277:DNA, Single-Stranded / Q000032:analysis*; D019075:Electrophoresis, Capillary / Q000295:instrumentation / Q000379:methods*; D005456:Fluorescent Dyes; D044085:Microfluidics / Q000295:instrumentation; D017422:Sequence Analysis, DNA / Q000379:methods*; D020080:Tandem Repeat Sequences; D000099166:Polyacrylamides
D000180:Acrylic Resins; D004277:DNA, Single-Stranded; D005456:Fluorescent Dyes; D000099166:Polyacrylamides