Pool-ARMS:一种高效检测混合遗传样本中特定单核苷酸变异的方法OACSTPCD

Single nucleotide variation(SNV)is one of the genetic basis for controlling human diseases and economic traits in animals and plants.An economical,simple,and high-efficient detection system is useful for screening genetic disease and screening plants carrying specific SNV in plant genomic editing and mutated populations.This study invented a method suitable for screening individuals carrying specific SNVs in rice pooled samples based on the amplification refractory mutation system(ARMS)PCR,which is referred as Pool-ARMS.The key steps of Pool-ARMS are as follow:Designing PCR primers and establishing an optimized PCR program for amplifying a fragment with specific SNV.Determining the amplification efficiency of pooled samples with DNA extracted from different tissues.Establishing a Pool-ARMS system for screening specific SNVs.Taking two genes that control photoperiod-sensitive genic male sterility(PGMS)in rice as examples,we first optimized the PCR primers and reaction program for amplifying of the two alleles,then we established leaf-and coleoptile-tissue based mutation detection systems with detection levels of 1∶49 and 1∶24,respectively.We genotyped 63 base edited rice seedlings for the targeted SNVs of PGMS using this Pool-ARMS system,with results consistent with Sanger sequencing analysis.This method is expected to be also useful for targeted screening of specific SNVs in pooled animal and plant genetic populations including rice materials.