METHOD FOR DETERMINING BASE SEQUENCE OF DNA OR DNA FRAGMENT BY AUTORADIOGRAPHY METHOD

摘要

PURPOSE:To obtain exactly positional information in short time by using an accelerated phosphor sheet by separating and developing a mixture composed of the synthesized DNA which has the base sequence complementary with the DNA to be determined in base sequence or the fragment thereof and is conjugated with a radioactive label. CONSTITUTION:Mixed DNA is synthesized by using >=1 kind of base labelled by <32>P, etc. in the stage of synthesizing complementary DNA by preparing the signal chain DNA (expressed as template DNA) of the DNA to be determined in the base sequence and hybridizing the complementary short DNA fragment (primer) to a part thereof. The mixed DNA is separated and developed by a gel electrophoresis method and thereafter a cumulative phosphor sheet contg. an accelerated phosphor is superposed on the separated and developed gel support for specified time then excitation light is scanned and irradiated to the phosphor sheet. The accelerated light emitted therefrom is detected and the positional information is obtd. in the form of a symbol or numerical value. The exact base sequence is thus determined in shorter time by the method using the X-ray film in the above-mentioned way.