Genetic screening of Leber's hereditary optic neuropathy by PCR with wholeblood cell lysate

Leber's hereditary optic neuropathy ( LHON) is accompanied by a mitochondri al DNA ( mtDNA) mutation. The G to A substitution at nucleotide position 11 ,778 (11,202) of mtDNA is most common in Japanese LHON patients. Whole bloo d cell lysate, not purified DNA, was used as a template of the polymerase c hain reaction ( PCR) for the analysis of the G11,778 (11,202) A point mutat ion. The amplified DNA fragment was concentrated and desalted with a centri fuge device, SUPREC(TM)-02, and digested by SfaNI and MaeIII. This method d oes not need purified DNA from blood and avoids the phenol/chloroform treat ments for PCR products prior to the restriction enzyme digestion. Therefore , it is convenient and safe for the genetic screening of LHON.