EXPANSION METHOD OF ARRAYED AMPLIFICATION OF RESTRICTION ENZYME FRAGMENTS

摘要

PURPOSE: An expansion method of arrayed amplification of restriction enzyme fragments is provided which expanses amplified total DNA or mRNA of single organism using PCR(polymerase chain reaction). CONSTITUTION: Using one of type IIs or type IIip restriction enzyme which generates nonspecific single stranded base sequence, total DNA or cDNA obtained from mRNA as a template of single organism is cleaved. The cleaved DNA fragments are ligated with adapters of DNA/RNA hybrid hairpin structure as shown in figure 1, and is treated with exonuclease to remove non-ligated DNA fragments. The ligated DNA fragments are treated with NaOH or RNase to remove RNA of the hairpin structure and to produce double stranded DNA. The obtained double stranded DNA is amplified by PCR.