Portable and Battery-Powered PCR Device for DNA Amplification and Fluorescence Detection.

Polymerase chain reaction (PCR) is a technique for nucleic acid amplification, which has been widely used in molecular biology. Owing to the limitations such as large size, high power consumption, and complicated operation, PCR is only used in hospitals or research institutions. To meet the requirements of portable applications, we developed a fast, battery-powered, portable device for PCR amplification and end-point detection.

Fluorescence quantification of intracellular materials at the single-cell level by an integrated dual-well array microfluidic device.

We present an integrated microfluidic device for quantifying intracellular materials at the single-cell level. This device combines a dual-well structure and a microfluidic control system. The dual-well structure includes capture wells (20 μm in diameter) for trapping a single cell and reaction wells (200 μm in diameter) for confining reagents.

One step DNA amplification of mammalian cells in picoliter microwell arrays.

We developed a strategy for direct DNA amplification of single cells on a PEG-modified silica chip with 30 600 picoliter-sized microwells. HPV-positive cells in heterogeneous populations were successfully detected with high accuracy sensitivity as high as single copy.

A novel dual-well array chip for efficiently trapping single-cell in large isolated micro-well without complicated accessory equipment.

Conventional cell-sized well arrays have advantages of high occupancy, simple operation, and low cost for capturing single-cells. However, they have insufficient space for including reagents required for cell treatment or analysis, which restricts the wide application of cell-sized well arrays as a single-cell research tool alone. Here, we present a novel dual-well array chip, which integrates capture-wells (20 m in diameter) with reaction-wells (100 m in diameter) and describe a flow method for convenient single-cell analysis requiring neither complicated infra-structure nor high expenditure, while enabling highly efficient single cell trapping (75.

Complete genome of Marinobacter psychrophilus strain 20041(T) isolated from sea-ice of the Canadian Basin.

Marinobacter psychrophilus strain 20041(T) was isolated from sea-ice of the Canadian Basin. Here we report the complete sequence of the 3.9-Mb genome of this strain.

Complete genome sequence of a deeply branched marine Bacteroidia bacterium Draconibacterium orientale type strain FH5(T).

Draconibacterium orientale strain FH5(T) isolated from a marine sediment sample from coast of Weihai, China, was a new species within the proposed new genus Draconibacterium in class Bacteroidia. Here, we present the genome sequence of D. orientale FH5(T), which contains 5,132,075 bp with a G+C content of 41.

Genome sequence and genetic diversity of the common carp, Cyprinus carpio.

The common carp, Cyprinus carpio, is one of the most important cyprinid species and globally accounts for 10% of freshwater aquaculture production. Here we present a draft genome of domesticated C. carpio (strain Songpu), whose current assembly contains 52,610 protein-coding genes and approximately 92.

Human pharyngeal microbiome may play a protective role in respiratory tract infections.

The human pharyngeal microbiome, which resides at the juncture of digestive and respiratory tracts, may have an active role in the prevention of respiratory tract infections, similar to the actions of the intestinal microbiome against enteric infections. Recent studies have demonstrated that the pharyngeal microbiome comprises an abundance of bacterial species that interacts with the local epithelial and immune cells, and together, they form a unique micro-ecological system. Most of the microbial species in microbiomes are obligate symbionts constantly adapting to their unique surroundings.

Heteroresistance at the single-cell level: adapting to antibiotic stress through a population-based strategy and growth-controlled interphenotypic coordination.

Unlabelled: Heteroresistance refers to phenotypic heterogeneity of microbial clonal populations under antibiotic stress, and it has been thought to be an allocation of a subset of "resistant" cells for surviving in higher concentrations of antibiotic. The assumption fits the so-called bet-hedging strategy, where a bacterial population "hedges" its "bet" on different phenotypes to be selected by unpredicted environment stresses. To test this hypothesis, we constructed a heteroresistance model by introducing a blaCTX-M-14 gene (coding for a cephalosporin hydrolase) into a sensitive Escherichia coli strain.

Improved picoliter-sized micro-reactors for high-throughput biological analysis.

High-throughput pyrosequencing, carried out in millions of picoliter-sized reactors on a fiber-optic slide, is known for its longer read length. However, both optical crosstalk (which reduces the signal-to-noise ratio of CCD images) and chemical retention adversely affect the accuracy of chemiluminescence determination, and ultimately decrease the read length and the accuracy of pyrosequencing results. In this study, both titanium and oxidized aluminum films were deposited on the side walls and upper faces of micro-reactor slides to enhance optical isolation; the films reduced the inter-well crosstalk by one order of magnitude.

Complete Genome Sequence of Marinobacter sp. BSs20148.

Marinobacter sp. BSs20148 was isolated from marine sediment collected from the Arctic Ocean at a water depth of 3,800 m. Here we report the complete genome sequence of Marinobacter sp.

Complete genome sequence of methicillin-sensitive Staphylococcus aureus containing a heterogeneic staphylococcal cassette chromosome element.

Staphylococcus aureus is a common human bacterium that sometimes becomes pathogenic, causing serious infections. A key feature of S. aureus is its ability to acquire resistance to antibiotics.

The history and advances of reversible terminators used in new generations of sequencing technology.

DNA sequencing using reversible terminators, as one sequencing by synthesis strategy, has garnered a great deal of interest due to its popular application in the second-generation high-throughput DNA sequencing technology. In this review, we provided its history of development, classification, and working mechanism of this technology. We also outlined the screening strategies for DNA polymerases to accommodate the reversible terminators as substrates during polymerization; particularly, we introduced the "REAP" method developed by us.

A pyrosequencing-based metagenomic study of methane-producing microbial community in solid-state biogas reactor.

Background: A solid-state anaerobic digestion method is used to produce biogas from various solid wastes in China but the efficiency of methane production requires constant improvement. The diversity and abundance of relevant microorganisms play important roles in methanogenesis of biomass. The next-generation high-throughput pyrosequencing platform (Roche/454 GS FLX Titanium) provides a powerful tool for the discovery of novel microbes within the biogas-generating microbial communities.

A complete genome assembly of Glaciecola mesophila sp. nov. sequenced by using BIGIS-4 sequencer system.

Using a pyrosequencing-based custom-made sequencer BIGIS-4, we sequenced a Gram-negative bacterium Glaciecola mesophila sp. nov. (Gmn) isolated from marine invertebrate specimens.

The next-generation sequencing technology and application.

As one of the key technologies in biomedical research, DNA sequencing has not only improved its productivity with an exponential growth rate but also been applied to new areas of application over the past few years. This is largely due to the advent of newer generations of sequencing platforms, offering ever-faster and cheaper ways to analyze sequences. In our previous review, we looked into technical characteristics of the next-generation sequencers and provided prospective insights into their future development.

[Nonsyndromic hereditary deafness genes research progress and related databases].

Deafness is the most prevalent sensory system impairment of human, and 70% of genetic deafness belongs to nonsyndromic hearing impairment. The total number of genes involved in nonsyndromic hereditary deafness has been estimated to above 100. So far, approximate 80 loci have been mapped to human chromosome, and 23 genes have been identified.

[Mutation analysis of Cx26 gene in Chinese hereditary nonsyndromic deafness sufferers].

Objective: To analyze the mutations in the code region of Cx26 gene in Chinese hereditary nonsyndromic hearing impairment (NSHI) sufferers.

Methods: Thirty-three cases (29 cases in the families of 8 students who were picked out from the Deafness and Muteness School of Tianjin, 2 cases as control and 2 normal cases of genetic counseling) were included in this study. The blood samples were obtained to distill the DNA templates.

[Restriction endonucleases fingerprinting-single strand conformation polymorphism: an efficient method to screen mutations in long segments].

Objective: To develop a simple, cheap and efficient restriction endonucleases fingerprinting-single strand conformation polymorphism(REF-SSCP) method applied to screen for mutations in long segments.

Methods: The genomic DNA of Cx26 gene segment of the patients with deafness was amplified. The amplification products were screened with SSCP and REF-SSCP technique and DNA sequencing to evaluate and compare the effect on detection of mutations in long segments.