1996
DOI: 10.1002/elps.1150170913
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High‐resolution capillary array electrophoretic sizing of multiplexed short tandem repeat loci using energy‐transfer fluorescent primers

Abstract: Short tandem repeat regions (STRs) from the polymorphic loci VWFA, THO1, TPO and CSF were amplified by the multiplex polymerase chain reaction (PCR) and analyzed by capillary array electrophoresis with fluorescence detection of energy transfer (ET) labels. The fluorescent ET primers are labeled with one fluorescein at the 5' end and a second fluorescein at the position of the 7th or 9th (modified) base to produce fragments that fluoresce in the green (lambda max = 525 nm). M13 A-track sequencing fragments, use… Show more

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Cited by 47 publications
(38 citation statements)
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“…for alleles 7-9 of 0.18 nucleotide, but a S.D. of 0.34 nucleotide for allele 6 (Wang et al 1996). By use of a similar polymer mix at room temperature, Mansfield et al (1996) report S.D.s for the THO1 allelic ladder fragments ranging from 0.10 to 0.14 nucleotide.…”
Section: Discussionmentioning
confidence: 98%
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“…for alleles 7-9 of 0.18 nucleotide, but a S.D. of 0.34 nucleotide for allele 6 (Wang et al 1996). By use of a similar polymer mix at room temperature, Mansfield et al (1996) report S.D.s for the THO1 allelic ladder fragments ranging from 0.10 to 0.14 nucleotide.…”
Section: Discussionmentioning
confidence: 98%
“…To increase the accuracy in sizing, it might be possible to calculate a normalizing factor (Mansfield et al 1996) or, alternatively, energy-transfer fluorescent dye-labeled DNA primers Wang et al 1995) that exhibit only very minor size differences could be used. More important than the absolute sizing accuracy, however, are highly reproducible analyses (Pritchard et al 1995;Wang et al 1996). These analyses should provide consistent genotyping for the same allelic STR, when using similar instrumentation and run conditions, independent from the genomic source and from run-to-run.…”
Section: Discussionmentioning
confidence: 99%
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“…The high throughput of these CAE microplates will facilitate rapid and inexpensive doublestranded restriction fragment screening for genetic diseases such as HHC. CAE microplates will also be valuable for single-stranded short tandem repeat analysis under denaturing conditions, following the methods recently developed by Wang and co-workers for forensic identification and cancer diagnosis (35,36).…”
Section: Discussionmentioning
confidence: 99%
“…To improve throughput, arrays of capillaries have been operated in parallel (14)(15)(16)). An analysis of the quadruplex STR system CTTv was reported on a 5-column array device with a throughput of 5 samples in 50 min (17). Also, a 48-column array system has been described where 48 samples could be analyzed in parallel in 1 h (18).…”
mentioning
confidence: 99%