Improved protocol for recovery of bacterial DNA from water filters: Sonication and backflushing of commercial syringe filters.
Extraction of bacterial DNA from water filters is frequently used in aquatic microbial ecology, e.g., for sequencing-based diversity studies. Here, we describe an improved size-class filtration and bacterial DNA extraction protocol using commercial “syringe filters”, involving sonication- and backflushing-steps and DNA-binding magnetic beads. Importantly, the sonication-step improved DNA recovery up to 10-fold. The DNA extraction-turnaround time is 4h, with a hands-on time of about 2h.
Fragmentation of DNA in a sub-microliter microfluidic sonication device.
Fragmentation of DNA is an essential step for many biological applications including the preparation of next-generation sequencing (NGS) libraries. As sequencing technologies push the limits towards single cell and single molecule resolution, it is of great interest to reduce the scale of this upstream fragmentation step.
Here we describe a miniaturized DNA shearing device capable of processing sub-microliter samples based on acoustic shearing within a microfluidic chip. A strong acoustic field was generated by a Langevin-type piezo transducer and coupled into the microfluidic channel via the flexural lamb wave mode.
Purified genomic DNA, as well as covalently cross-linked chromatin were sheared into various fragment sizes ranging from ∼180 bp to 4 kb. With the use of standard PDMS soft lithography, our approach should facilitate the integration of additional microfluidic modules and ultimately allow miniaturized NGS workflows
Fragmentation of DNA by sonication.
INTRODUCTIONDNA fragmentation is often necessary prior to library construction or subcloning for DNA sequencing. This protocol describes a method for DNA fragmentation by sonication. During sonication, DNA samples are subjected to hydrodynamic shearing by exposure to brief periods of sonication.
DNA that has been sonicated for excessive periods of time is extremely difficult to clone. Most sonicators will not shear DNA to a size of less than 300-500 bp, and it is tempting to continue sonication until the entire DNA population has been reduced in size. However, the yield of subclones is usually greater if sonication is stopped when the fragments of the target DNA first reach a size of ~700 bp.
Description: Anti-Flag magnetic beads is based on hydroxyl magnetic beads covalently coupling with mouse IgG1 monoclonal antibody. It is recommended to use for co-immunoprecipitation and protein purification.
Description: Anti-Flag magnetic beads is based on hydroxyl magnetic beads covalently coupling with mouse IgG1 monoclonal antibody. It is recommended to use for co-immunoprecipitation and protein purification.
Description: Anti-HA magnetic beads kit is based on hydroxyl magnetic beads covalently coupling with high quality mouse IgG2b monoclonal antibody, used for co-immunoprecipitation and protein purification.
Description: Anti-HA magnetic beads kit is based on hydroxyl magnetic beads covalently coupling with high quality mouse IgG2b monoclonal antibody, used for co-immunoprecipitation and protein purification.
Description: Anti-Myc magnetic beads kit is based on hydroxyl magnetic beads covalently coupling with mouse IgG1 monoclonal antibody.it is recommended to use for co-immunoprecipitation and protein purification.
Description: Anti-Myc magnetic beads kit is based on hydroxyl magnetic beads covalently coupling with mouse IgG1 monoclonal antibody.it is recommended to use for co-immunoprecipitation and protein purification.
Description: Protein A/G Magnetic Beads for immunoprecipitation (IP) are based on a biological nanosurface technology (S-TEC). Protein A/G coats the surface of super paramagnetic microspheres with high coating density up to 9.3 × 10^13 molecules/cm2. Compared to similar immune magnetic beads, our Protein A/G beads display much more antibody binding sites therefore using less magnetic beads per reaction and ultimately achiving greater cost-efficiency. Non-specific binding is reduced to an absolute low eliminating any detectable interference with your IP assays. The large, specific surface area of the beads greatly shortens the adsorption time and achieves complete antibody/antigen adsorption process in 10 minutes and complete total purification and precipitation in just 30 minutes. This product can be used on a wide variety of samples, such as in cell lysates, supernatants collected from cell secretion, serum, ascites, and other immune antigens.
Description: Protein A/G Magnetic Beads for immunoprecipitation (IP) are based on a biological nanosurface technology (S-TEC). Protein A/G coats the surface of super paramagnetic microspheres with high coating density up to 9.3 × 10^13 molecules/cm2. Compared to similar immune magnetic beads, our Protein A/G beads display much more antibody binding sites therefore using less magnetic beads per reaction and ultimately achiving greater cost-efficiency. Non-specific binding is reduced to an absolute low eliminating any detectable interference with your IP assays. The large, specific surface area of the beads greatly shortens the adsorption time and achieves complete antibody/antigen adsorption process in 10 minutes and complete total purification and precipitation in just 30 minutes. This product can be used on a wide variety of samples, such as in cell lysates, supernatants collected from cell secretion, serum, ascites, and other immune antigens.
Description: Amino Magnetic Particles are prepared by coating a layer of magnetite and polystyrene onto monodispersed (i.e.. uniform sized) polystyrene core particles. The magnetic particles can be easily separated from a suspension magnetically. These particles become non magnetic when removed from a magnet, and do not retain any detectable magnetism even after repeated exposure to strong magnetic field.Are of research that those particles had been tested are cell separation, affinity purification, DNA probe assays, magnetic particle EIA. For more infomation please don't hesiate to contact our technical team.
Description: Please reffer to the technical data sheet for more detail information for this item. Our dedicated team would be happy to assist you via live chat, email or phone.
Description: Please reffer to the technical data sheet for more detail information for this item. Our dedicated team would be happy to assist you via live chat, email or phone.
Description: Please reffer to the technical data sheet for more detail information for this item. Our dedicated team would be happy to assist you via live chat, email or phone.
Description: Please reffer to the technical data sheet for more detail information for this item. Our dedicated team would be happy to assist you via live chat, email or phone.
Description: Please reffer to the technical data sheet for more detail information for this item. Our dedicated team would be happy to assist you via live chat, email or phone.
Description: Please reffer to the technical data sheet for more detail information for this item. Our dedicated team would be happy to assist you via live chat, email or phone.
Description: Please reffer to the technical data sheet for more detail information for this item. Our dedicated team would be happy to assist you via live chat, email or phone.
Description: HIF pathway activator (EC50 values are 1.23 and 1.4 ?M for HRE gene reporter assay and HIF-1? nuclear translocation assay respectively); acts via chelation of iron, independently of PHD.
Description: ML239 is the best-in-class inhibitor of the breast cancer stem cells with an IC50= 1.16 µM. [1]ML239 was a selective inhibitor an IC50= 1.18 µM against HMLE_sh_Ecad.
Description: ML-356 is an inhibitor of the thioesterase domain of fatty acid synthase with IC50 value of 334 nM.Fatty acid synthase (FAS) is a multi-enzyme protein that catalyzes fatty acid synthesis, fatty acids are related to energy production and storage, cellular structure ,hormones biosynthesis
Description: IC50: 32, 20, and 41 nM for TC-83, V3526, and Trinidad donkey strains, respectively.ML-336 is an inhibitor for VEEV-induced cytopathic effect.
Description: IC50: LYPLA1 (17 nM) and the related LYPLA2 (30 nM)ML-211 is a dual inhibitor of LYPLA1 and the related LYPLA2.Lysophospholipase 1 (LYPLA1), a protein palmitoyl thioesterase, is responsible for depalmitoylation of the oncogene HRas.
Chromatin immunoprecipitation (ChIP): revisiting the efficacy of sample preparation, sonication, quantification of sheared DNA, and analysis via PCR.
The “quantitative” ChIP, a tool commonly used to study protein-DNA interactions in cells and tissue, is a difficult assay often plagued with technical error. We present, herein, the process required to merge multiple protocols into a quick, reliable and easy method and an approach to accurately quantify ChIP DNA prior to performing PCR.
We demonstrate that high intensity sonication for at least 30 min is required for full cellular disruption and maximum DNA recovery because ChIP lysis buffers fail to lyse formaldehyde-fixed cells. In addition, extracting ChIP DNA with chelex-100 yields samples that are too dilute for evaluation of shearing efficiency or quantification via nanospectrophotometry.
However, DNA extracted from the Mock-ChIP supernatant via the phenol-chloroform-isoamyl alcohol (PCIA)method can be used to evaluate DNA shearing efficiency and used as the standard in a fluorescence-based microplate assay.
This enabled accurate quantification of DNA in chelex-extracted ChIP samples and normalization to total DNA concentration prior to performing real-time PCR (rtPCR). Thus, a quick ChIP assay that can be completed in nine bench hours over two days has been validated along with a rapid, accurate and repeatable way to quantify ChIP DNA.
The resulting rtPCR data more accurately depicts treatment effects on protein-DNA interactions of interest.
Rapid quantification methods for genetically modified maize contents using genomic DNAs pretreated by sonication and restriction endonuclease digestion for a capillary-type real-time PCR system with a plasmid reference standard.
For rough quantitative analysis of genetically modified maize contents, rapid methods for measurement of the copy numbers of the cauliflower mosaic virus 35S promoter region (P35S) and MON810 construct-specific gene (MON810) using a combination of a capillary-type real-time PCR system with a plasmid DNA were established.
To reduce the characteristic differences between the plasmid DNA and genomic DNA, we showed that pretreatment of the extracted genomic DNA by a combination of sonication and restriction endonuclease digestion before measurement is effective.
The accuracy and reproducibility of this method for MON810 content (%) at a level of 5.0% MON810 mixed samples were within a range from 4.26 to 5.11% in the P35S copy number quantification. These methods should prove to be a useful tool to roughly quantify GM maize content.
Description: A competitive ELISA for quantitative measurement of Rabbit Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Rabbit Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Rabbit Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Goat Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Goat Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Goat Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Dog Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
Description: A competitive ELISA for quantitative measurement of Canine Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Dog Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
Description: A competitive ELISA for quantitative measurement of Canine Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Dog Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
Description: A competitive ELISA for quantitative measurement of Canine Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Porcine Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Porcine Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Porcine Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Mouse Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Mouse Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Mouse Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Rat Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
Description: A competitive ELISA for quantitative measurement of Rat Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Rat Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
Description: A competitive ELISA for quantitative measurement of Rat Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Rat Rhizopus arrhizu Fatty Acid Desaturases 6 ELISA kit
Description: A competitive ELISA for quantitative measurement of Rat Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Monkey Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Monkey Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Monkey Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Guinea pig Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Guinea pig Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Guinea pig Rhizopus arrhizu Fatty Acid Desaturases 6 in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.