METHOD FOR DETERMINATION OF TRANSGENIC LINE GA21 OF MAIZE BY MEANS OF POLYMEARSE CHAIN REACTION

摘要

A method for detection of mutant gene of 5-enolpiruvil shikimate-3-phosphatesynthase of maize (Zea mays L.) in genetically modified plant by means of polymerase chain reaction, to this end the thermal denaturation of plant DNA is made; cyclic amplification, where each cycle comprises the denaturation of DNA, renaturation of plant DNA with oligonucleotide primers, synthesis of fragments of target genes. For polymerase chain reaction the pairs of oligonucleotide primers are used, complementary to mutant gene of 5-enolpiruvil shikimate-3-phosphatesynthase of maize, flanking genome DNA. The subsequent amplification is made under the following conditions: denaturation of plant DNA during 4 min. at 94 °C; 35 cycles, each of them comprises the denaturation of DNA during 30 sec. at 94 °C, renaturation of plant DNA with oligonucleotide primers during 30 sec. at 55 °C, synthesis of fragments of target genes during 15 sec. at 72 °C; final synthesis of fragments of target genes during 10 min. at 72 °C.