Detection Of Minimal Residual Leukemia Cells In Acute Myeloid Leukemia By Polymerase Chain Reaction Single Strand Conformational Polymorphism

Polymerase chain reaction (PCR) is a method for the primer -directed enzymatic amplification of specific DNA sequences in vitro and continuation of PCR for a few hours may result in 10~7-10~8times amplification of DNA region of interest. Single strand conformational polymorphism (SSCP) is an elec-trophoresis technique of polyacrylamide gels, by which single stranded DNA or RNA fragments, even only one base is different , can be seperated in gels according to the difference of folded structure of nucleic acids.The patients with acute myeloid leukemia (AMD were screened for N-ras gene mutations and followed by using PCR -SSCP.First, we determined the sensitivity of the PCR-SSCP using two-step PCR-SSCP technique.HL-60 cell line DNA,which possess gene mutations in the second position of codon 61 of N-ras, were diluted with normal DNA. After the first PCR which was non-radioisotopic and symmetric and SSCP were performed, the gels, in which mutational single strands located, were cut out, and then mutational single strands were isolated from gels by electrophoresis. The second PCR which was radioisoto-...