Due to their high specificity and affinity towards various targets,along with other unique advantages such as stability and low cost,aptamers are widely applied in analytical techniques.A typical aptamerbased electroc...Due to their high specificity and affinity towards various targets,along with other unique advantages such as stability and low cost,aptamers are widely applied in analytical techniques.A typical aptamerbased electrochemical biosensor is composed of a aptamer as the biological recognition element and transducer converting the biologic interaction into electrical signals for the quantitative measurement of targets.Improvement of the sensitivity of a biosensor is significantly important in order to achieve the detection of biomolecules with low abundance,and different amplification strategies have been explored.The strategies either employ nanomaterials such as gold nanoparticles to con struct electrodes which can transfer the biological reactions more efficiently,or attempt to obtain enhanced signal through multi-labeled carriers or utilize enzyme mimics to catalyze redox cycling.This review discusses recent advances in signal amplification methods and their applications.Critical assessment of each method is also considered.展开更多
The extraction of nucleic acid is recognized as one of the most essential steps in molecular biology for initiating other downstream applications such as sequencing, amplification, hybridization, and cloning. Many com...The extraction of nucleic acid is recognized as one of the most essential steps in molecular biology for initiating other downstream applications such as sequencing, amplification, hybridization, and cloning. Many commercial kits and methods are currently available that allow the extraction of only one type of nucleic acids-DNA or RNA. However, in parallel clinical detection of several diseases, a method for simultaneous extraction of both DNA and RNA from the same source is needed in such cases. In this study, a method for simultaneous extraction of DNA and RNA from bacteria based on magnetic nanoparticles(MNPs) was described. Lysis buffers were prepared to help the nucleic acid released and adsorbed to MNPs. Then, two washing buffers were used to remove the contamination of proteins and carbohydrates. The nucleic acids were finally eluted by Deoxyribonuclease(DNase) and Ribonucleases(RNase) free water. Different factors which might affect the purification of the nucleic acid were investigated, and the quantity and quality parameters of the nucleic acid were also recorded. The DNA and RNA extracted from bacteria were then respectively subjected to polymerase chain reaction(PCR) and reverse transcription PCR(RT-PCR) to further confirm its quality. The results indicated that our method can be successfully used to simultaneously extract DNA and RNA from bacteria.展开更多
基金This work was financially supported by the National Key Research and Development Program of China(No.2017YFA0205301)the National Natural Science Foundation of China(Nos.61527806,81902153 and 61871180)+1 种基金the Clinical Advanced Technology of Social Development Projects in Jiangsu Province(No.BE2018695)the Natural Science Foundation of Hunan Province(No.2017JJ2069).
文摘Due to their high specificity and affinity towards various targets,along with other unique advantages such as stability and low cost,aptamers are widely applied in analytical techniques.A typical aptamerbased electrochemical biosensor is composed of a aptamer as the biological recognition element and transducer converting the biologic interaction into electrical signals for the quantitative measurement of targets.Improvement of the sensitivity of a biosensor is significantly important in order to achieve the detection of biomolecules with low abundance,and different amplification strategies have been explored.The strategies either employ nanomaterials such as gold nanoparticles to con struct electrodes which can transfer the biological reactions more efficiently,or attempt to obtain enhanced signal through multi-labeled carriers or utilize enzyme mimics to catalyze redox cycling.This review discusses recent advances in signal amplification methods and their applications.Critical assessment of each method is also considered.
基金supported by the National Basic Research Program of China(2014CB744501)the National High Technology Research and Development Program of China(2012AA022703)+8 种基金the National Key Special Science Program(2013ZX10004103-002)the National Natural Science Foundation of China(61201033,21205013,61271056,61527806)Projects of Development of Science and Medical Technology(201208038)Projects of Health Ministry of Nanjing(ZKX12038)the Clinical Science and Technology Special Projects in Jiangsu Province(BL2012067,BL2014094)the Talents Planning of Six Summit Fields of Jiangsu Province(2013-WSN-056)China Postdoctoral Science Foundation Funded Project(2014M551491,2015T80487)Jiangsu Planned Projects for Postdoctoral Research Funds(1302007A)the Economical Forest Cultivation and Utilization of 2011 Collaborative Innovation Center in Hunan Province
文摘The extraction of nucleic acid is recognized as one of the most essential steps in molecular biology for initiating other downstream applications such as sequencing, amplification, hybridization, and cloning. Many commercial kits and methods are currently available that allow the extraction of only one type of nucleic acids-DNA or RNA. However, in parallel clinical detection of several diseases, a method for simultaneous extraction of both DNA and RNA from the same source is needed in such cases. In this study, a method for simultaneous extraction of DNA and RNA from bacteria based on magnetic nanoparticles(MNPs) was described. Lysis buffers were prepared to help the nucleic acid released and adsorbed to MNPs. Then, two washing buffers were used to remove the contamination of proteins and carbohydrates. The nucleic acids were finally eluted by Deoxyribonuclease(DNase) and Ribonucleases(RNase) free water. Different factors which might affect the purification of the nucleic acid were investigated, and the quantity and quality parameters of the nucleic acid were also recorded. The DNA and RNA extracted from bacteria were then respectively subjected to polymerase chain reaction(PCR) and reverse transcription PCR(RT-PCR) to further confirm its quality. The results indicated that our method can be successfully used to simultaneously extract DNA and RNA from bacteria.
