Four ClEF1A genes involved in self-incompatibility in 'Xiangshui Lemon' confer early fowering and increase stress tolerance in transgenic Arabidopsis.

The plant elongation factor eEF1A is involved in coregulating not only the translation of proteins and controlling translation-related signaling but also in signaling associated with cell growth, stress response and motility, controlling apoptosis and responding to adversity in plants. In this study, four eEF1A genes, namely, ClEF1A-1, ClEF1A-2, ClEF1A-3 and ClEF1A-4, were identified from the genomic and ubiquitin-modified omics data of the 'Xiangshui Lemon', and bioinformatics analysis revealed that these four genes have relatively similar structures with conserved sequences; ClEF1A-1 and ClEF1A-4 were highly expressed in pollen, and temporal expression analysis demonstrated that the expression of ClEF1As was significantly greater under self-pollination than under cross-pollination. All four genes were localized in the nucleus.

Abridged solid-phase extraction with alkaline Poly(ethylene) glycol lysis (ASAP) for direct DNA amplification.

Complexity of sample preparation decelerate the development of sample-in-answer-out devices for point-of-need nucleic acid amplification testing. Here, we present the consolidation of alkaline poly(ethylene) glycol-based lysis and solid-phase extraction for rapid and simple sample preparation compatible with direct on-bead amplification. Simultaneous cell lysis and binding of DNA were achieved using an optimised reagent comprising 15% PEG8000, 0.

Assessing the use of environmental DNA (eDNA) as a tool in the detection of human DNA in water.

Environmental DNA (eDNA) is a highly sensitive and cost-effective tool that is increasingly being applied to studies of biodiversity and species detection. This non-invasive method relies on the collection of environmental samples that contain genetic material being shed into surrounding environment by the target organism/s. While forensic science has a long history of using molecular tools for collecting DNA from the environment, the detection of human DNA from environmental water samples has been limited.

An improved nucleic acid sequence-based amplification method mediated by T4 gene 32 protein.

The uptake of Nucleic Acid Sequence-Based Amplification (NASBA) for point of care testing may be hindered by a complexity in the workflow due the requirement of a thermal denaturation step to initiate the cyclic isothermal amplification before the addition of the amplification enzymes. Despite reports of successful enhancement of other DNA and RNA amplification methods using DNA and RNA binding proteins, this has not been reported for NASBA. Here, three single-stranded binding proteins, RecA, Extreme Thermostable Single-stranded binding protein (ET SSB) and T4 gene gp32 protein (gp32), were incorporated in NASBA protocol and used for single pot, one-step NASBA at 41 °C.

Isotachophoresis for rapid transformation of Escherichia coli.

A frequent limitation of electroporation (EP) and chemical transformation (CT) are the need of tedious and time-consuming procedures for inducing transformation competence, the substantial number of cells required, and the low transformation yields typically achieved. Here, we show a new and rapid electrokinetic method for transformation of small number of noncompetent Escherichia coli TOP10 cells (2-3 × 10 ) at room temperature. Escherichia coli TOP10 cells and plasmid DNA are sequentially injected into a 50 μm ID capillary and focused into 11.

Detecting marine pests using environmental DNA and biophysical models.

The spread of marine pests is occurring at record rates due to globalisation and increasing trade. Environmental DNA (eDNA) is an emerging tool for pest surveillance, allowing for the detection of genetic material shed by organisms into the environment. However, factors influencing the spatial and temporal detection limits of eDNA in marine environments are poorly understood.

Application of Microfluidic Methodology for the Analysis of DNA.

Over the past 20 years, many of the developments and potential applications of microfluidic methodology have incorporated nucleic acid processes which have, in their own right, undergone a number of innovative changes [...

Counter-pressure-assisted ITP with electrokinetic injection under field-amplified conditions for bacterial analysis.

Counter-pressure was used to extend the duration of field-amplified sample injection in isotachophoresis (FASI-ITP) in order to improve the detection of bacterial cells. Using 0.51-μm negatively charged encapsulated fluorescent beads as a model, the counter-pressure, injection and separation voltages, and times were optimized.

Polymeric microchip for the simultaneous determination of anions and cations by hydrodynamic injection using a dual-channel sequential injection microchip electrophoresis system.

A dual-channel sequential injection microchip capillary electrophoresis system with pressure-driven injection is demonstrated for simultaneous separations of anions and cations from a single sample. The poly(methyl methacrylate) (PMMA) microchips feature integral in-plane contactless conductivity detection electrodes. A novel, hydrodynamic "split-injection" method utilizes background electrolyte (BGE) sheathing to gate the sample flows, while control over the injection volume is achieved by balancing hydrodynamic resistances using external hydrodynamic resistors.

Sieving polymer synthesis by reversible addition fragmentation chain transfer polymerization.

Replaceable sieving polymers are the fundamental component for high resolution nucleic acids separation in CE. The choice of polymer and its physical properties play significant roles in influencing separation performance. Recently, reversible addition fragmentation chain transfer (RAFT) polymerization has been shown to be a versatile polymerization technique capable of yielding well defined polymers previously unattainable by conventional free radical polymerization.

Rapid and sensitive microbial analysis by capillary isotachophoresis with continuous electrokinetic injection under field amplified conditions.

A highly sensitive capillary isotachophoresis method with LIF detection for microbial analysis was developed. This allowed the reliable analysis of Escherichia coli bacteria with a LOD of 14 cells in a sample volume of 100 μL, or 1.35 × 10(2) cell/mL, which is 47 times lower than reported by CE-LIF and 148 times lower than CE-UV with on-line concentration.

Capillary electrophoretic system of ribonucleic acid molecules.

Over the past two decades, capillary electrophoresis (CE) has been the subject of extensive development and progress in various DNA based sieving electrophoresis applications, namely Sanger sequencing, forensic short tandem repeat (STR) analysis, clinical genotype screening (SNP), and phylogenetic fingerprinting. Yet, this trend has not been emulated in the RNA field. This review will highlight the development and key analysis parameters of RNA electrophoresis by CE and provide possible explanations for the low research interest in this area.

Capillary electrophoresis ribosomal RNA single-stranded conformation polymorphism: a new approach for characterization of low-diversity microbial communities.

Capillary electrophoresis (CE) has been the principle system for nucleic acid analysis since the early 1990s due to its inherent advantages such as fast analysis time, high resolution and efficiency, minimal sample requirement, high detection sensitivity, and automation. In the past few decades, microbial community fingerprinting methods such as terminal restriction fragment length polymorphism and single-stranded conformation polymorphism (SSCP) have migrated to CE to utilize its advantages over conventional slab gel electrophoresis. Recently, a gel-based direct rRNA fingerprint method was demonstrated.

Identification of inorganic improvised explosive devices using sequential injection capillary electrophoresis and contactless conductivity detection.

A simple sequential injection capillary electrophoresis (SI-CE) instrument with capacitively coupled contactless conductivity detection (C(4)D) has been developed for the rapid separation of anions relevant to the identification of inorganic improvised explosive devices (IEDs). Four of the most common explosive tracer ions, nitrate, perchlorate, chlorate, and azide, and the most common background ions, chloride, sulfate, thiocyanate, fluoride, phosphate, and carbonate, were chosen for investigation. Using a separation electrolyte comprising 50 mM tris(hydroxymethyl)aminomethane, 50 mM cyclohexyl-2-aminoethanesulfonic acid, pH 8.