Ws. Oetting et al., Multiplexed short tandem repeat polymorphisms of the Weber 8A set of markers using tailed primers and infrared fluorescence detection, ELECTROPHOR, 19(18), 1998, pp. 3079-3083
Short tandem repeat polymorphism (STRP) markers have become important reage
nts for mapping genetic diseases. These markers are available as screening
sets, which are located in all chromosomes at discrete intervals, allowing
the entire genome to be analyzed. Mapping studies that include many individ
uals in the analysis necessitate the production of large numbers of genotyp
es. In an effort to increase the efficiency and lower the cost of using the
se STRP screening sets, we have divided the amplification primers of the We
ber 8A screening set into groups that can be amplified in single polymerase
chain reaction (PCR) amplification reactions, resulting in a reduction of
both time and cost. Fluorescently-labeled amplification products were produ
ced using a three primer reaction. The forward STRP amplification primer fo
r each marker contained a 19 bp sequence at the 5' end. A fluorescently-lab
eled primer, with a sequence identical to the 19 bp tail, was added to the
amplification reaction as the sole source of fluorescent label. The STRP ba
nding pattern is detected using an automated fluorescent DNA sequencer. Use
of this multiplexed genomic screening set should greatly enhance the mappi
ng of human disease loci.