title: Fast amplification of the low density lipoprotein receptor gene and detection of a large deletion by means of long polymerase chain reaction.
abstract: To demonstrate the usefulness of Long PCR in analyzing gene structures and large deletions we have developed a method to amplify the entire LDL receptor gene, including the promoter region and intron 1. This opens new ways for studies of the gene and allows the detection of certain LDL receptor-specific deletions. For the amplification of the LDL receptor gene, spanning approximately 45.5 x 10(3) bases and divided into 18 exons and 17 introns, we have designed overlapping PCR products (ranging from 4 to 16 x 10(3) bases in length), which can be amplified simultaneously overnight for fast results. It was possible to positively identify two samples from heterozygote carriers of the "5 kB French-Canadian" deletion using this method. As a side result the length of intron 1 of the LDL receptor gene could be established to be approximately 9.5 x 10(3) bases. The method is sensitive enough to detect deletions in 1:10 mixes of positive control with wildtype DNA.
authors.affiliation: Institut für Klinische Chemie und Laboratoriumsmedizin, Universitätsklinikum, Technischen Universität Dresden, Freistaat Sachsen, Germany.
authors.forename: H
authors.initials: H
authors.lastname: Bochmann
authors.forename: S
authors.initials: S
authors.lastname: Gehrisch
authors.forename: W
authors.initials: W
authors.lastname: Jaross
mesh_terms: D017353:Gene Deletion; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D011973:Receptors, LDL / Q000235:genetics*
chemical_list: D011973:Receptors, LDL