Automation of genetic linkage analysis using fluorescent microsatellite markers.
Automation of the typing of genetic markers offers advantages of speed, accuracy, and cost in the mapping of genetic traits and the construction of high-resolution linkage maps. We have developed an automated linkage analysis system by (i) using a robotic pipettor to set up polymerase chain reactions (PCR) to amplify microsatellites with incorporation of a single fluorescent label; (ii) using an automated sequencing apparatus for detection of the PCR products; (iii) sizing alleles automatically by the use of internal and external standards; (iv) iteratively filtering out nonallelic fragments and checking for Mendelian consistency; (v) calculating the probabilities of selected genotypes; and (vi) automatically formatting the results for input to linkage analysis programs. The method provides accurate sizing of alleles, minimizes the risk of error during manual reading and transcription of data, and increases the throughput of reliable data. It brings any inconsistencies or ambiguities in the data to the attention of the user and facilitates examination of the raw data. The ALF/ALP system, together with new, optimized microsatellite sets, particularly tetranucleotide repeats, is likely to be well-suited to fully automatic genetic linkage analysis.
Mansfield
D C
DC
MRC Human Genetics Unit, Western General Hospital, Edinburgh, United Kingdom.
Brown
A F
AF
Green
D K
DK
Carothers
A D
AD
Morris
S W
SW
Evans
H J
HJ
Wright
A F
AF
D000483:Alleles; D001331:Automation; D004276:DNA, Satellite / Q000235:genetics*; D005456:Fluorescent Dyes; D008040:Genetic Linkage; D005819:Genetic Markers; D005821:Genetic Techniques; D005838:Genotype; D016133:Polymerase Chain Reaction
D004276:DNA, Satellite; D005456:Fluorescent Dyes; D005819:Genetic Markers