| 摘要 |
A method for detection of transformation event GT73 in rape in genetically modified plant by multiplex polymerase chain reaction includes denaturation of plant DNA; cycles each containing denaturation of DNA, renaturation of plant DNA with oligonucleotide primers, synthesis of target genes fragments. For carrying out of polymerase chain reaction pairs of oligonucleotide primers are used, specific to transformation event GT73 (1 μM GT1F; 1 μM GT1R), to the gene CP4 epsps, responsible for plant resistance towards glyphosate (0.25 μM 4EPSPSF; 0.25 μM 4EPSPSR), and pair of primers to fragment of own rape gene acetyl-coenzyme A carboxylase (0.75 μM acc2; 0.75 μM acc3), and amplification is carried out at the following conditions: denaturation of plant DNA for 4 minutes under 94°C; 34 cycles, each comprising denaturation of DNA for 30 sec under 94 °C, renaturation of plant DNA with oligonucleotide primers for 30 sec under 62 °C, synthesis of target genes fragments for 30 sec under 72 °C, synthesis of target genes fragments for 10 minutes under 72 °C. |
