The persistence length and the overlap concentration(c~*) of poly(ethylene oxide)(PEO) and hydroxyethylcellulose(HEC) with similar molecular weight in 1×TBE buffer were studied by laser light scattering and visco...The persistence length and the overlap concentration(c~*) of poly(ethylene oxide)(PEO) and hydroxyethylcellulose(HEC) with similar molecular weight in 1×TBE buffer were studied by laser light scattering and viscometry.Their effect on DNA separation was investigated by capillary electrophoresis.It was determined that the persistence length of HEC was at least 5 times higher than that of PEO.Therefore,the c~* of HEC was smaller than that of PEO by a factor of ca.2.5.It was also found that the c~* values deter...展开更多
The mixtures of two polymers, poly (N,N-dimethylacrylamide) (PDMA) and polyvinylpyrrolidone (PVP) were synthesized and used as the separation medium for double-stranded and single-stranded DNA fragments by capillary e...The mixtures of two polymers, poly (N,N-dimethylacrylamide) (PDMA) and polyvinylpyrrolidone (PVP) were synthesized and used as the separation medium for double-stranded and single-stranded DNA fragments by capillary electrophoresis with UV detector. On optimal conditions, 2%w/v PDMA + 2%w/v PVP can be used to separate the doublet 123/124bp in pBR322/Hae III Markers.展开更多
Pyricularia oryzae is a fungal pathogen of rice.Few researches had been reported on the analysisof its genome, partly because the genomes can’t beseparated by general agarose gel electrophoresis.We herein report a te...Pyricularia oryzae is a fungal pathogen of rice.Few researches had been reported on the analysisof its genome, partly because the genomes can’t beseparated by general agarose gel electrophoresis.We herein report a technique to separatePyricularia oryzae genomic DNA by pulse-fieldgel electrophoresis(PFGE).The tested blast strain was 2539w.Chromosomal DNAs were directly prepared from展开更多
A PDMS electrophoresis microchip,which integrated with optical fiber for fluorescence detection,was fabricated by using silicon master.A deep reactive ion etches (DRIE) technology was used to fabricate the silicon ma...A PDMS electrophoresis microchip,which integrated with optical fiber for fluorescence detection,was fabricated by using silicon master.A deep reactive ion etches (DRIE) technology was used to fabricate the silicon master with positive features.The PDMS replica was fabricated by casting PDMS prepolymer against the silicon master,where an optical fiber was first fixed on the end of separation microchannel.To improve the rigid characteristics of integrated PDMS microchip,the chips were subsequently assembled by reversible sealing against glass plate.A blue light emitting diode (LED) was used as excitation light sources for inducing fluorescence detection through coupling LED light into the optical fiber.As an application, integrated PDMS microchip was tested in the capillary electrophoresis separation of DNA markers.The results showed that DNA markers could be effectively separated and detected except for the segments of 271 and 281.展开更多
We have previously developed bare narrow-bore capillary chromatography. In this work, high-performance DNA separation was realized for a size range of 10–800 base pairs(bp) utilizing bare narrow-bore capillary chroma...We have previously developed bare narrow-bore capillary chromatography. In this work, high-performance DNA separation was realized for a size range of 10–800 base pairs(bp) utilizing bare narrow-bore capillary chromatography with 750 nm- radius capillaries. Separation behavior of double-stranded DNA(ds DNA) fragments was investigated over a range of eluent concentrations and elution pressures. DNA molecules were hydrodynamically separated in a size-dependent manner in free solution without any sieving matrices, with the longer fragments being eluted out from the capillary earlier. It was found that the eluent concentration variously influenced the transport behavior for different-sized DNA fragments depending upon the configuration of DNA molecules and the association of counterions. Ionic strength of the solutions strongly impacted DNA persistence length. Enhanced elution pressure could shorten analysis time with a slight loss in resolution. Excellent efficiency of two million theoretical plates per meter was achieved, which indicates the enormous potential of bare narrow-bore capillary chromatography for the analysis of DNA fragments. These findings would be useful in understanding the transport behavior of DNA fragments in confined dimensions for chromatography in free solution.展开更多
A new approach is presented for preparative,continuous flow fractionation of sub-10-kbp DNA fragments,which exploits the variation in the field-dependent mobility of the DNA molecules based on their length.Orthogonall...A new approach is presented for preparative,continuous flow fractionation of sub-10-kbp DNA fragments,which exploits the variation in the field-dependent mobility of the DNA molecules based on their length.Orthogonally pulsed electric fields of significantly different magnitudes are applied to a microchip filled with a sieving matrix of 1.2% agarose gel.Using this method,we demonstrate a high-resolution separation of 0.5,1,2,5,and 10 kbp DNA fragments within 2 min.During the separation,DNA fragments are also purified from other ionic species.Preparative fractionation of sub-10-kbp DNA molecules plays an important role in second-generation sequencing.The presented device performs rapid high-resolution fractionation and it can be reliably manufactured with simple microfabrication procedures.展开更多
基金supported by the National Natural Science Foundation of China(No20504001)
文摘The persistence length and the overlap concentration(c~*) of poly(ethylene oxide)(PEO) and hydroxyethylcellulose(HEC) with similar molecular weight in 1×TBE buffer were studied by laser light scattering and viscometry.Their effect on DNA separation was investigated by capillary electrophoresis.It was determined that the persistence length of HEC was at least 5 times higher than that of PEO.Therefore,the c~* of HEC was smaller than that of PEO by a factor of ca.2.5.It was also found that the c~* values deter...
基金supported by the Innovation Fund(KSCX1-06)of Chinese Academy of Sciences.
文摘The mixtures of two polymers, poly (N,N-dimethylacrylamide) (PDMA) and polyvinylpyrrolidone (PVP) were synthesized and used as the separation medium for double-stranded and single-stranded DNA fragments by capillary electrophoresis with UV detector. On optimal conditions, 2%w/v PDMA + 2%w/v PVP can be used to separate the doublet 123/124bp in pBR322/Hae III Markers.
文摘Pyricularia oryzae is a fungal pathogen of rice.Few researches had been reported on the analysisof its genome, partly because the genomes can’t beseparated by general agarose gel electrophoresis.We herein report a technique to separatePyricularia oryzae genomic DNA by pulse-fieldgel electrophoresis(PFGE).The tested blast strain was 2539w.Chromosomal DNAs were directly prepared from
基金This present work was supported by grant from the National Natural Science Foundation of China (N0．60501020， No．60341005 and No．20299030)
文摘A PDMS electrophoresis microchip,which integrated with optical fiber for fluorescence detection,was fabricated by using silicon master.A deep reactive ion etches (DRIE) technology was used to fabricate the silicon master with positive features.The PDMS replica was fabricated by casting PDMS prepolymer against the silicon master,where an optical fiber was first fixed on the end of separation microchannel.To improve the rigid characteristics of integrated PDMS microchip,the chips were subsequently assembled by reversible sealing against glass plate.A blue light emitting diode (LED) was used as excitation light sources for inducing fluorescence detection through coupling LED light into the optical fiber.As an application, integrated PDMS microchip was tested in the capillary electrophoresis separation of DNA markers.The results showed that DNA markers could be effectively separated and detected except for the segments of 271 and 281.
基金supported by the National Natural Science Foundation of China(21275014)the Excellent Young Scientists Fund of NSFC(21322501)+3 种基金the Importation and Development of High-Caliber Talents Project of Beijing Municipal Institutions(CIT&TCD20140309)the Program for New Century Excellent Talents in University(NCET-12-0603)the Beijing Natural Science Foundation ProgramScientific Research Key Program of Beijing Municipal Commission of Education(KZ201310005001)
文摘We have previously developed bare narrow-bore capillary chromatography. In this work, high-performance DNA separation was realized for a size range of 10–800 base pairs(bp) utilizing bare narrow-bore capillary chromatography with 750 nm- radius capillaries. Separation behavior of double-stranded DNA(ds DNA) fragments was investigated over a range of eluent concentrations and elution pressures. DNA molecules were hydrodynamically separated in a size-dependent manner in free solution without any sieving matrices, with the longer fragments being eluted out from the capillary earlier. It was found that the eluent concentration variously influenced the transport behavior for different-sized DNA fragments depending upon the configuration of DNA molecules and the association of counterions. Ionic strength of the solutions strongly impacted DNA persistence length. Enhanced elution pressure could shorten analysis time with a slight loss in resolution. Excellent efficiency of two million theoretical plates per meter was achieved, which indicates the enormous potential of bare narrow-bore capillary chromatography for the analysis of DNA fragments. These findings would be useful in understanding the transport behavior of DNA fragments in confined dimensions for chromatography in free solution.
文摘A new approach is presented for preparative,continuous flow fractionation of sub-10-kbp DNA fragments,which exploits the variation in the field-dependent mobility of the DNA molecules based on their length.Orthogonally pulsed electric fields of significantly different magnitudes are applied to a microchip filled with a sieving matrix of 1.2% agarose gel.Using this method,we demonstrate a high-resolution separation of 0.5,1,2,5,and 10 kbp DNA fragments within 2 min.During the separation,DNA fragments are also purified from other ionic species.Preparative fractionation of sub-10-kbp DNA molecules plays an important role in second-generation sequencing.The presented device performs rapid high-resolution fractionation and it can be reliably manufactured with simple microfabrication procedures.
