Event-specific qualitative PCR detection method of stress-resistant soybean IND-??41?-5 and its standardization
Stress-resistant soybean IND-ØØ41Ø-5,which contains HaHB4 gene and bar marker gene,has been granted a safety certificate for importation as a processing material into China.In this study,We designed 19 primer pairs based on the 5'and 3'flanking sequences of the stress-resistant soybean IND-ØØ41Ø-5 event,in combination with one pair of primer providing by Instituto de Agrobiotecnologia Rosario(INDEAR)S.A.The specif-ic screening for the 20 primer pairs was performed using qualitative PCR technology.The results indicated that the primer pair of 14 at the 5'flanking sequence had good specificity.After optimization,the best reaction system and reaction procedure were obtained.The primer pair was tested for their specificity and stability,with limit of detec-tion(LOD)of 0.1%and an amplified fragment size of 248 bp.The specificity,LOD,repeatability and reproducibility of this method were verified by eight qualified laboratories,all of which met the requirements of national standards.This study successfully established a qualitative PCR method to detect the stress-resistant soybean IND-ØØ41Ø-5 event,which can provide technical support for the safety regulation of IND-ØØ41Ø-5 event in China.
万方数据
维普
