Fast amplification of the low density lipoprotein receptor gene and detection of a large deletion by means of long polymerase chain reaction. Eur J Clin Chem Clin Biochem

Summary: 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 Χ 103 bases and divided into 18 exons and 17 introns, we have designed overlapping PCR products (ranging from 4 to 16 Χ 103 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 Χ 103 bases. The method is sensitive enough to detect deletions in 1: 10 mixes of positive control with wildtype DNA