Analysis of multiplexed short tandem repeat (STR) systems using capillary array electrophoresis.
The profiling of polymorphic short tandem repeat (STR) markers is being applied to human identification, parentage testing and genetic mapping. Reliable genotyping of these markers is facilitated by polymerase chain reaction (PCR) amplification and high-resolution electrophoretic separation. Capillary array electrophoresis (CAE) offers very rapid, high-resolution separation of the amplified DNA and potential for automated sample processing not realized employing conventional slab-gel electrophoresis. The use of CAE to type DNA samples amplified at 11 genetic loci in multiplex profiles is presented. Two sets totaling 208 samples were amplified in a multiplex fashion using AmpFlSTR-Blue or AmpFlSTR-Green I and analyzed in a blind study using CAE. With the exception of one sample, the CAE genotyping results were in complete agreement with results obtained using a single-capillary system or two slab-gel electrophoresis systems. The sample, genotype TH01 7/10, migrated similar to TH01 6.3/9.3 allele sizes, which suggested a potential band migration shift. The recommended approach to such an observation is to analyze the sample again. The sample was rerun and correct genotype verified. Allelic ladder samples were analyzed multiple times by CAE to determine sizing accuracy and precision. The sizing of over 240 allelic ladder samples yielded an average within-run precision of +/- 0.13 bp and between-run precision of +/- 0.21 bp for fragments up to 350 bp. The CAE protocols permit processing of up to 96 multiplex STR samples in under 70 min.
Mansfield
E S
ES
Molecular Dynamics, Inc., Sunnyvale, CA 94086, USA. emansfield@mdyn.com
Robertson
J M
JM
Vainer
M
M
Isenberg
A R
AR
Frazier
R R
RR
Ferguson
K
K
Chow
S
S
Harris
D W
DW
Barker
D L
DL
Gill
P D
PD
Budowle
B
B
McCord
B R
BR
D000483:Alleles; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D019075:Electrophoresis, Capillary / Q000379:methods*; D005838:Genotype; D006801:Humans; D012091:Repetitive Sequences, Nucleic Acid; D015203:Reproducibility of Results
D004247:DNA