Improved PCR Amplification/Hhal Restriction for Unambiguous Determination of Apolipoprotein E

We present a modification to the polymerase chain reaction amplification/Hhal restriction isotyping method for human apolipoprotein (apo) E. This method includes a mutagenic forward primer and 5’-end labeling of both primers. These modifications of the original method de-scribed by Hixon and Vernier(J Lipid Res 1990;31:545-8) allow sensitive and unambiguous determination of apoE genotypes. Indexing Terms: genotype determination/polymerase chain mac-tion/restriction endonucleases Apolipoprotein E (apoE) is an important constituent of several classes of plasma lipoproteins, predomi-nantly very-low-density and high-density lipoproteins and chylomicrons.2 ApoE is a ligand of the receptor for low-density lipoprotein and of the putative apoE (chy