Objective The detection of RNA single nucleotide polymorphism(SNP)is of great importance due to their association with protein expression related to various diseases and drug responses.At present,splintR ligase-assist...Objective The detection of RNA single nucleotide polymorphism(SNP)is of great importance due to their association with protein expression related to various diseases and drug responses.At present,splintR ligase-assisted methods are important approaches for RNA direct detection,but its specificity will be limited when the fidelity of ligases is not ideal.The aim of this study was to create a method to improve the specificity of splintR ligase for RNA detection.Methods In this study,a dualcompetitive-padlock-probe(DCPLP)assay without the need for additional enzymes or reactions is proposed to improve specificity of splintR ligase ligation.To verify the method,we employed dual competitive padlock probe-mediated rolling circle amplification(DCPLP-RCA)to genotype the CYP2C9 gene.Results The specificity was well improved through the competition and strand displacement of dual padlock probe,with an 83.26%reduction in nonspecific signal.By detecting synthetic RNA samples,the method demonstrated a dynamic detection range of 10 pmol/L-1 nmol/L.Furthermore,clinical samples were applied to the method to evaluate its performance,and the genotyping results were consistent with those obtained using the qPCR method.Conclusion This study has successfully established a highly specific direct RNA SNP detection method,and provided a novel avenue for accurate identification of various types of RNAs.展开更多
As an important wild blueberry resource,Vaccinium uliginosum has attracted more and more attention.At present,the wild resources are under destruction.The conservation of wild Vaccinium uliginosum resources is imminen...As an important wild blueberry resource,Vaccinium uliginosum has attracted more and more attention.At present,the wild resources are under destruction.The conservation of wild Vaccinium uliginosum resources is imminent.However,there are few researches on the protection and preservation of its germplasm resources.In vitro preservation is an important method for germplasm conservation.In this study,one strain of wild Vaccinium uliginosum was used as material.The effects of temperature(25℃,15℃,10℃,or 0℃),media(WPM,1/2WPM or 1/3WPM),medium supplements(sorbitol or mannose),and photoperiod(8,10,12,or 14 h•d^(-1))on the growth,survival rate and rejuvenation rate of the plantlets were studied.The physiological changes of plantlets during preservation were analyzed.Methylation-sensitive amplified polymorphism(MSAP)analysis of genomic DNA methylation of plantlets was carried out to explore the genetic stability of the plantlets after preservation.The research results provided a theoretical basis for the germplasm preservation of Vaccinium uliginosum.展开更多
文摘Objective The detection of RNA single nucleotide polymorphism(SNP)is of great importance due to their association with protein expression related to various diseases and drug responses.At present,splintR ligase-assisted methods are important approaches for RNA direct detection,but its specificity will be limited when the fidelity of ligases is not ideal.The aim of this study was to create a method to improve the specificity of splintR ligase for RNA detection.Methods In this study,a dualcompetitive-padlock-probe(DCPLP)assay without the need for additional enzymes or reactions is proposed to improve specificity of splintR ligase ligation.To verify the method,we employed dual competitive padlock probe-mediated rolling circle amplification(DCPLP-RCA)to genotype the CYP2C9 gene.Results The specificity was well improved through the competition and strand displacement of dual padlock probe,with an 83.26%reduction in nonspecific signal.By detecting synthetic RNA samples,the method demonstrated a dynamic detection range of 10 pmol/L-1 nmol/L.Furthermore,clinical samples were applied to the method to evaluate its performance,and the genotyping results were consistent with those obtained using the qPCR method.Conclusion This study has successfully established a highly specific direct RNA SNP detection method,and provided a novel avenue for accurate identification of various types of RNAs.
基金Supported by the National Natural Science Foundation of China(32172521)。
文摘As an important wild blueberry resource,Vaccinium uliginosum has attracted more and more attention.At present,the wild resources are under destruction.The conservation of wild Vaccinium uliginosum resources is imminent.However,there are few researches on the protection and preservation of its germplasm resources.In vitro preservation is an important method for germplasm conservation.In this study,one strain of wild Vaccinium uliginosum was used as material.The effects of temperature(25℃,15℃,10℃,or 0℃),media(WPM,1/2WPM or 1/3WPM),medium supplements(sorbitol or mannose),and photoperiod(8,10,12,or 14 h•d^(-1))on the growth,survival rate and rejuvenation rate of the plantlets were studied.The physiological changes of plantlets during preservation were analyzed.Methylation-sensitive amplified polymorphism(MSAP)analysis of genomic DNA methylation of plantlets was carried out to explore the genetic stability of the plantlets after preservation.The research results provided a theoretical basis for the germplasm preservation of Vaccinium uliginosum.
