Identification of novel simple sequence length polymorphisms (SSLPs) in mouse by interspersed repetitive element (IRE)-PCR.
Interspersed repetitive element (IRE)-PCR is a useful method for identification of novel human or mouse sequence tagged sites (STSs) from contigs of genomic clones. We describe the use of IRE-PCR with mouse B1 repetitive element primers to generate novel, PCR amplifiable, simple sequence length polymorphisms (SSLPs) from yeast artificial chromosome (YAC) clones containing regions of mouse chromosomes 13 and 14. Forty-two IRE-PCR products were cloned and sequenced from eight YACs. Of these, 29 clones contained multiple simple sequence repeat units. PCR analysis with primers derived from unique sequences flanking the simple sequence repeat units in seven clones showed all to be polymorphic between various mouse strains. This novel approach to SSLP identification represents an efficient method for saturating a genomic interval with polymorphic genetic markers that may expedite the positional cloning of genes for traits and diseases.
Detter
J C
JC
Department of Medicine and Center for Mammalian Genetics, University of Florida, 1600 SW Archer Road,PO Box 100221, Gainesville, FL 32610, USA. jdetter@nervm.nerdc.ufl.edu
Nguyen
Q A
QA
Kingsmore
S F
SF
D000818:Animals; D001483:Base Sequence; D018244:Chromosomes, Artificial, Yeast; D017931:DNA Primers; D051379:Mice; D008815:Mice, Inbred Strains / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D012091:Repetitive Sequences, Nucleic Acid; D016324:Sequence Tagged Sites; D013045:Species Specificity
D017931:DNA Primers