The lipoprotein receptor-associated protein (RAP) is considered a chaperone protein for the lipoprotein receptor-related protein (LRP) and for the other members of the LDL receptor family. Genetic analysis is anticipated to help in delineating groups or individuals with potential defects or problems in this regard. A combined amplification/sequencing strategy was developed to analyze the human LRPAP1 gene for polymorphisms and mutations. The LRPAP1 gene was amplified from genomic DNA in four long-range PCR amplicons, 2.4 to 7.6 kb in size. Three amplicons were finally used as templates with 14 sequencing primers to obtain the sequence of the eight exons and large portions of adjacent introns from individual DNA. This strategy, applied to sequence the LRPAP1 gene of 14 unrelated, normal individuals revealed, in total, 23 distinct mutations and polymorphisms, mostly intronic substitutions and deletions. In this small group 1 expressed mutation was encountered on one allele in 2 unrelated individuals: a G to A transition results in the replacement of valine by methionine in exon 7 at position 311 of the human RAP precursor protein.