... ,High Molecular Weight DNA Markers are suitable for sizing linear double-stranded DNA from 9 to 48 kb on low-percentage agarose gels. The 13 bands consist of 8.3- to 48.5-kb fragments generated from restriction endonuclease digests of Lambda DNA (cIind1ts857 Sam7). Ethidium bromide staining causes t,biological,biology supply,biology supplies,biology product
Ammonium sulfate protein precipitation is a key technique for concentration, fractionation and purification of proteins. Salting-out is a key technique.
As depicted in Fig. 1, acidic phospholipids such as PIP3 (derived from conversion of PIP2 to PIP3 by PI3K, e.g., as occurring during insulin action) and PA (derived from PLD action on various phospholipids or from de novo phospholipid synthesis) most likely bind to undefined basic arginine residues in or near the pseudosubstrate site and thereby activate the enzyme by promoting 1) molecular unfolding, 2) dissociation of the pseudosubstrate sequence from the substrate-binding site, and 3) opening of the critically needed phosphorylation sites at Thr403 and Thr555 (25, 47). PKCι can also be phosphorylated on tyrosine residues 256, 271, and 325, which may account for the direct activation of aPKC by Src kinase (58). AMPK, which activates glucose transport via aPKC (see below), also directly phosphorylates purified PKCζ, and it was postulated that AMPK directly phosphorylates Thr410, the activation loop site in PKCζ, presumably independently of PDK1, and this leads to activation (18). However, ...
Two novel members of the human cAMP-dependent protein kinase inhibitor (PKI) gene family, PKIB and PKIG, were cloned. The deduced proteins showed 70% and 90% identity with mouse PKIβ and PKIγ respectively. Both the already identified pseudosubstrate site and leucine-rich nuclear export signal motifs were defined from the 11 PKIs of different species. The PKIB and PKIG genes were mapped respectively to chromosome 6q21-22.1, using a radiation hybrid GB4 panel, and to chromosome 20q13.12-13.13, using a Stanford G3 panel. Northern-blot analysis of three PKI isoforms, including the PKIA identified previously, revealed significant differences in their expression patterns. PKIB had two transcripts of 1.9 kb and 1.4 kb. The former transcript was abundant in both placenta and brain and the latter was expressed most abundantly in placenta, highly in brain, heart, liver, pancreas, moderately in kidney, skeletal muscle and colon, and very little in the other eight tissues tested. PKIG was widely expressed ...
However, in the case of PCR generated templates, Ambion adds amplified DNA directly to the transcription reaction with no purification. 5 μl of a 100 μl PCR reaction (or about 0.05 - 0.2 μg of double-stranded DNA) is used as template. However, with shorter templates or low yields, the concentration of template in a 5 μl aliquot of the crude PCR reaction may be suboptimal. In that case, it may be desirable to concentrate the PCR product by alcohol precipitation. We do not generally find it necessary to phenol/chloroform extract the PCR reaction before precipitation, although in some cases it may be advisable to do so. [1 ...
The GenElute HP Plasmid Miniprep Kits offer purification of high-quality plasmid DNA in less than 30 minutes. These kits provide the convenience of binding columns that can be used with a vacuum or spin purification format. In addition, no alcohol precipitation is required.
Im wondering if the Hirt high molecular weight DNA prep has been replaced by a simpler or kit like method. I have to isolate a viral DNA from some infected tissue culture cells and would like to make it as painless as possible. Thanks for your help. Mary ...
In recent years Alcohol precipitation has been declining and one of the processes that has taken its place is the gel press or KCl precipitation process. This technology comes from agar processing and is essentially identical to the manner in which agar is made from gracillaria type weed stocks. Eucheuma Cottonii has a very high kappa carrageenan content and when purified by filtration it is then possible to precipitate the carrageenan using potassium chloride, generally this forms a large jelly like mass that can be pressed to remove water. The pressed cake can then be dried and milled as usual. The great advantage of this process is that you have produced a refined product without incurring the extra costs of the alcohol recovery. On the downside the process only works with Eucheuma Cottonii and can only make kappa type carrageenan gels. This process has had great succession the large Asian dessert jellies market.. There are two processes that aim to cheapen the production of carrageenan by ...
Rx Biosciences Glycogen is a highly purified polysaccharide derived from oysters. It is most widely used as an inert carrier which significantly increases the recovery of nucleic acids by alcohol precipitation. Being insoluble in ethanol, Glycogen forms a precipitate that traps target nucleic acids. During centrifugation, a visible pellet is formed, which greatly facilitates handling of the precipitated nucleic acids. Glycogen quantitatively precipitates nucleic acids from diluted solutions with a higher efficiency than that of tRNA, linear polyacrylamide, or sonicated DNA.. ...
We are not concerned about mixed samples, because there is no preferential amplification with REPLI-g WGA. A small level of contamination from a secondary source cannot become a major component of the amplified material ...
My laboratory has achieved excellent incorporation of 32P following the protocol in Rubin, G. 1976. Methods in Cell Biology 12:45-64. The phosphate depleted media described in that paper is made by selective precipitation of inorganic phosophate (as MgNH4PO4) from YEPD. On Tue, 12 Sep 2000, Brian Baker wrote: , Im having a hard time trying to label my yeast cells with 32P. the , label does not seem to be incorporated. Im using 32P in the form of , H3PO4, Im growing my cells in -phosphate media, and I label them for , ~45 min at 30 C. Does anyone know whats I can do? or have a better , protocol? , , --- ______________O_________oO_____________oO______o_______oO___________________ YEAST bionet newsgroup see: http://www.bio.net/hypermail/YEAST/ YEAST e-mail: messages sent to yeast at net.bio.net subscribe: e-mail biosci-server at net.bio.net with: subscribe yeast unsubscribe: e-mail biosci-server at net.bio.net with: unsubscribe yeast YEAST on the WWW: ...
Analysis of Extracted DNA Samples Samples of DNA were quantitatively assayed using spectrophotometery by measuring absorbance at 260 nm (A260), 280 nm (A280), and 230 nm (A230) to determine concentration of ds DNA, and to measure A260 to A280 ratio (optimal ratio is equal to 1.8 or more and less than 2) and A260 to A230 ratio (optimal ratio is greater than 2.2). A280 is an estimate of concentration of non-DNA components. such as proteins, whereas A230 is measured to estimate concentration of other components such as detergents, peptides, carbohydrates, Inhibitors,research,lifescience,medical phenol and chloroform. Some randomly selected samples were assayed quantitatively by running on a 2% agarose gel electrophoresis, Inhibitors,research,lifescience,medical allowing the molecular weight of the DNA to be estimated. The electrophoresis resulted in a thick smear of DNA fragments with variable size ranging from very high molecular weight DNA to those about 100 bp in size (figure 1). Moreover, some ...
The association between telomere length (TL) in peripheral blood cells and cancer risk remains inconclusive. We carried out a meta-analysis on prospective studies. The study-specific RR estimates were first transformed to a common comparable scale and then were pooled by a random-effects model. The dataset was composed of 13,894 cases and 71,672 controls from 28 studies in 25 articles. In the comparison of the longest versus shortest third of TL, we observed a marginally positive association between longer TL and higher risk of total cancers [OR = 1.086; 95% confidence interval (CI), 0.952-1.238]. Subgroup analyses showed that the association was stronger in lung cancer (n = 3; OR = 1.690; 95% CI, 1.253-2.280), in men (n = 6; OR = 1.302; 95% CI, 1.120-1.514) and in studies with more precise methods for DNA extraction (phenol-chloroform, salting-out or magnetic bead, n = 6, OR = 1.618; 95% CI, 1.320-1.985) and TL measurement (multiplex Q-PCR, n = 8; OR = 1.439; 95% CI, 1.118-1.852). Our ...
Selective precipitation of gold from aqua regia, leaving pgm in solution february 25, 2010 am an analytical chemist for a recycler am new to this area and have a question regarding a selective precipitation of gold from an anolyte solution of 10 by volume aqua regia.
SUMMARY: The enzymes of N-acetyl-D-glucosamine (GlcNAc) metabolism, GlcNAc-6-phosphate deacetylase and GlcN-6-phosphate deaminase were found to be inducible in Candida albicans. The pattern of induction for these enzymes was the same under conditions of germ-tube formation (37 °C) and where yeast cells metabolized GlcNAc with no change in morphology (28 °C); this indicates that these enzymes are not control points in the dimorphic development of C. albicans. During induction there was a 40- and 25-fold increase in specific activity for the deacetylase and the deaminase, respectively, and the maximum specific activity corresponded to the time when all the GlcNAc had been metabolized. The presence of lomofungin (an inhibitor of transcription) or trichodermin (an inhibitor of translation) in cell suspensions of C. albicans containing GlcNAc prevented the increase in specific activity of these enzymes. 2-Deoxyglucose inhibited germ-tube formation, partially inhibited the induction of the deacetylase (43%)
The mechanism of binding of immune complexes to macrophages was investigated using purified antibody and haptens of different valences. Antibody alone bound to macrophages; enhancement of binding occurred when polyvalent and divalent haptens were present at equivalence but did not occur in great antigen excess. Monovalent hapten did not increase the binding of antibody at any concentration ratio tried, though it inhibited the enhancement due to oligovalent hapten. Ultracentrifuged normal rabbit globulin also inhibited the binding of complexes indicating the presence of exposed binding sites on the uncomplexed molecules. Complexes bound more strongly than antibody alone as determined from elution studies. These results support the hypothesis that the enhancement of antibody binding to macrophages in the presence of antigen is due to increased energy of binding resulting from summation of individual binding sites already exposed on the antibody molecules.. It was also possible, by saturating the ...
Hyalin is a protein released from the cortical granules of a fertilized animal egg. The released hyalin modifies the extracellular matrix of the fertilized egg to block other sperm from binding to the egg, and is known as the slow-block to polyspermy. All animals have this slow-block mechanism. Hyalin is a large, acidic protein which aids in embryonic development. The protein has strong adhesive properties which can help with cell differentiation and as a polyspermy prevention component. It forms the hyaline layer which covers the surface of the egg after insemination. Its physical structure has a major and minor component. One is filamentous, having flexible molecules containing a globular domain head at the end. Its conformation is retained mainly by disulfide bonds, as virtually all cysteine amino acids are found in the disulfide form, but also hydrophobic forces and salt linkages stabilize the molecule. The filament length is about 75 nm long, and the head being club-shaped with a diameter ...
1Environmental Microbial Genomics Group, Laboratoire AMPERE, Ecole Centrale de Lyon, Université de Lyon, 36 avenue Guy de Collongue, 69134 Ecully, France. Metagenomic studies require obtaining relatively high quantities of DNA and/or high molecular weight DNA from complex environments. Furthermore, if the interest is only in procaryotic DNA, contamination by eucaryotic DNA may represent an important issue. These technical constraints depend on the type of metagenomic approach: whereas high throughput sequencing (e.g. pyrosequencing) requires several micrograms of DNA without strict size constraints, the construction of fosmid or cosmid clone libraries requires relatively lower amounts of DNA but whose size must be , 25kb. We focused here on two technical aspects: (i) obtaining procaryotic DNA free of eucaryotic contamination, and (ii) obtaining high quantities of DNA. To avoid contamination by eucaryotic DNA, an indirect extraction method is used, requiring a preliminary extraction of bacterial ...
1Environmental Microbial Genomics Group, Laboratoire AMPERE, Ecole Centrale de Lyon, Université de Lyon, 36 avenue Guy de Collongue, 69134 Ecully, France. Metagenomic studies require obtaining relatively high quantities of DNA and/or high molecular weight DNA from complex environments. Furthermore, if the interest is only in procaryotic DNA, contamination by eucaryotic DNA may represent an important issue. These technical constraints depend on the type of metagenomic approach: whereas high throughput sequencing (e.g. pyrosequencing) requires several micrograms of DNA without strict size constraints, the construction of fosmid or cosmid clone libraries requires relatively lower amounts of DNA but whose size must be , 25kb. We focused here on two technical aspects: (i) obtaining procaryotic DNA free of eucaryotic contamination, and (ii) obtaining high quantities of DNA. To avoid contamination by eucaryotic DNA, an indirect extraction method is used, requiring a preliminary extraction of bacterial ...
Read "Polythermal study of the salting-out of triethylamine from aqueous solutions with sodium nitrate, Russian Journal of Applied Chemistry" on DeepDyve, the largest online rental service for scholarly research with thousands of academic publications available at your fingertips.
Use the ReadyPrep 2-D cleanup kit to free protein samples of ionic contaminants by selective precipitation. Remove detergents, lipids, and phenolic compounds.
We have studied the posttranslational modifications of the 52-kD protein, an estrogen-regulated autocrine mitogen secreted by several human breast cancer cells in culture (Westley, B., and H. Rochefort, 1980, Cell, 20:353-362). The secreted 52-kD protein was found to be phosphorylated mostly (94%) on high-mannose N-linked oligosaccharide chains, and mannose-6-phosphate signals were identified. The phosphate signal was totally removed by alkaline phosphatase hydrolysis. The secreted 52-kD protein was partly taken up by MCF7 cells via mannose-6-phosphate receptors and processed into 48- and 34-kD protein moieties as with lysosomal hydrolases. By electron microscopy, immunoperoxidase staining revealed most of the reactive proteins in lysosomes. After complete purification by immunoaffinity chromatography, we identified both the secreted 52-kD protein and its processed cellular forms as aspartic and acidic proteinases specifically inhibited by pepstatin. The 52-kD protease is secreted in breast ...
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Article The implement of the electro-static and magnetic precipitation technique through the centeral air-conditioning systems. In this paper we will introduce the electrostatic and magnetic precipitation technique, to deposit the solid dust within t...
The N-end rule pathway is a proteolytic system where its recognition components (N-recognins) recognize destabilizing N-terminal residues of short-lived proteins as an important part of specific degrons called N-degrons. like a scaffold E3 that promotes HR6B/UbcH2-reliant ubiquitylation of H2A and H2B however not H3 and H4 via a system distinct from normal polyubiquitylation. The E3 activity of UBR2 in histone ubiquitylation is activated by dipeptides bearing destabilizing N-terminal residues allosterically. Insufficient monoubiquitylation and polyubiquitylation on UBR2-lacking meiotic chromosomes correlate to problems in dual strand break (DSB) restoration along with other meiotic procedures leading to pachytene arrest at stage IV and apoptosis. A few of these features of UBR2 are found in somatic cells where UBR2 is really a chromatin-binding proteins involved in chromatin-associated ubiquitylation upon DNA damage. UBR2-deficient somatic cells show an array of chromosomal abnormalities ...
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Press Release issued Oct 12, 2017: Future Market Insights (FMI) delivers key insights on the global and Far East & Baltic nations ammonium sulphate market in its upcoming report titled, Ammonium Sulphate Market: Global Industry Analysis and Opportunity Assessment, 2015-2025. The global ammonium sulphate market is projected to expand at a CAGR of 4.0% in terms of value during the forecast period owing to various factors, regarding which FMI offers vital insights in detail in this report.
Dr. bob could you please check this work cus i cant seem to get it right the first step of the synthesis is described by the reaction below. When 1.750 g of Fe(NH4)2(SO4)2 6H2O is mixed with 13 mL of 1.0 M H2C2O4, the theoretical ...
Recently, investigators at Universität Ulm have been investigating the low molecular weight DNA diffusion assay (LMW assay) to measure cytotoxicity in comet assays.. The comet assay is a well-established in vitro and in vivo genotoxicity test. However, there has always been concern that the comet assay (as with all DNA strand break assays) may possibly detect exposure-related cytotoxicity rather than genotoxic effects. This is because DNA degradation is frequently involved in processes leading to cell death. Concurrent measures for cytotoxicity are required for standard genotoxicity testing to assess genotoxicity and to address possible effects of cytotoxicity in comet assay data interpretation.. Typically, histopathology has been accepted as a suitable cytotoxicity detection method when using the in vivo comet assay. By looking at the histology, levels of necrosis and apoptosis can be determined. However, there are other assays available for assessing cytotoxicity.. Here, the LMW assay has ...
The effect of insulin administration on immune complex (IC) formation in diabetic patients was analysed in vivo and in vitro. Firstly, serial studies of IC status were performed over a mean period of 18 months in 44 diabetic patients, 37 of whom were receiving standard insulin therapy. Thirty patients changed to monocomponent (MC) insulin while seven commenced MC insulin after tablet failure. The other seven patients remained on standard insulin throughout the study. Secondly, nine patients had serial measurements of IC over a 6-8 h period following a routine morning dose of MC insulin; eight control subjects were similarly studied. The insulin content of IC in insulin treated patients was assessed in vitro by examining, (a) the selective precipitation of antibody bound insulin by 3% polyethylene glycol (PEG) and (b) the insulin specificity of antisera raised against PEG precipitates of IC positive sera. The longitudinal study of circulating IC showed no significant changes apart from an isolated fall
Moniezia a genus of tapeworms that are parasitic in mammals, including sheep, goat and cattle. It comprises four known species such as M. expansa, M. benedeni, M. autumnalis and M. baeri. M. expansa is the most well known species within the genus because of its high prevalence. Members of the genus are among the largest cestodes reaching up to 10 m in length. They inhabit the small intestine of mammalian host. Their life cycle is indirect requiring intermediate host, which are oribatid mites. They are characterized by the presence of interproglottid glands. Mehlhorn H (2008). Encyclopedia of parasitology, Volume 1 (3rd edn). Springer. ISBN 978-3-540-48994-8 BioLib ZipcodeZoo The ...
Morphological variants of intrahepatocytic hyalin in Indian childhood cirrhosis have been analysed by electron microscopy. This structure, morphologically identical with Mallorys alcoholic hyalin, is encountered in three different forms. The most common one is composed of randomly oriented fibrils. The next common type is composed of smudgy homogeneous or finely granular material, while the least common type consists of parallel fibrils with gentle curves giving a finger-print appearance. Fragmented endoplasmic reticulum and ribosomes were frequently associated with hyalin suggesting that these organelles contribute to their formation.. ...
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Precipitation in FBS - posted in Tissue and Cell Culture: Dear Guys, I have a new bottle of CCB FBS it is 500 ml bottle I have aliquot in 50 ml bottles. At first, there are 20 ml extra, this mean it was 520 ml not 500 only. so is it normal There are precipitation in a bottle of my FBS?? Any suggestion shall I filter my FBS and continue??? or shall I discard the FBS bottle with ppt
We evaluated and optimized three rapid methods for extraction of high-quality DNA from carbonate-encrusted microbial communities using modern calcifying oncolites built by cyanobacteria and diatoms in a high-calcium freshwater river. Pulverization, acid (HCl) dissolution, and chelator-mediated (EDTA) dissolution of the carbonate matrix were used and optimized to liberate microbial cells from their mineral encasing. This was followed by cell lysis and DNA extraction and isolation. HCl dissolution yielded no measurable DNA, probably due to hydrolysis, whereas pulverization and EDTA dissolution yielded averages of 3.5 and 7.8 μg per gram of sample, respectively, of high molecular weight DNA. These DNA isolates could be used successfully for PCR-amplification of 16S rRNA gene segments (alleles) and subsequent fingerprinting of the cyanobacterial (including diatoms) and total bacterial communities through denaturing gradient gel electrophoresis (DGGE) separation. Fingerprints showed no differences
Circulomics, a Baltimore, MD-based biotechnology startup, received two SBIR grants totaling over $1.7M. The company will use the funds to develop new applications of its Nanobind DNA/RNA extraction technology.. These new awards bring total funding to nearly $8M. Previous NIH grants have been used to develop kits for high molecular weight DNA extraction from cultured cells, bacteria, blood, and tissues.. The first award is a Phase I grant from the National Human Genome Research Institute (NHGRI) to develop Nanobind methods for tunable size selection purification in next-generation sequencing library preparation. Nanobind can be used for reaction cleanup to remove small background molecules such as adapters and primer dimers. However, Nanobinds unique binding mechanism also allows it to perform tunable size selection of much larger library products to enhance sequencing read lengths. All Nanobind methods use a simple bind, wash, and elute process that features high purification and high recovery ...
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Eurosolids Ammonium Sulphate Horticultural Grade is a highly pure Nitrogen fertilizer which dissolve rapidly and completely. Our product is not sensitive to caking and is recommended during periods of vegetative growth. The high level of Nitrogen in our formula improves cell elongation and contributes to the production of protein, supporting enzymatic reactions. ...
This clear 96-well filter plate has a Hydrophobic PTFE membrane used for Peptide-bead cleavage & cleanup, in-plate protein precipitation, sample recovery following in-plate compound cleavage, peptide synthesis. Find MSDS or SDS, a COA, data sheets and more information.
Protein precipitation technique is commonly used with biological samples. Study results show Bond Elut EMR-Lipid selectively removes lipids without affecting analyte recovery.
As an online gathering place for the fertilizer industry, Fertilizerworks.com serves manufacturers of agricultural nutrients and equipment; suppliers of distribution and application services; farmers; and other members of the fertilizer community. All are people and organizations who help grow the worlds food. ...
TY - JOUR. T1 - First insight into the genome of an uncultivated crenarchaeote from soil. AU - Quaiser, Achim. AU - Ochsenreiter, Torsten. AU - Klenk, Hans-Peter. AU - Kletzin, Arnulf. AU - Treusch, Alexander H. AU - Meurer, Guido. AU - Eck, Jürgen. AU - Sensen, Christoph W. AU - Schleper, Christa. PY - 2002. Y1 - 2002. N2 - Molecular phylogenetic surveys based on the characterization of 16S rRNA genes have revealed that soil is an environment particularly rich in microbial diversity. A clade of crenarchaeota (archaea) has frequently been detected among many other novel lineages of uncultivated bacteria. In this study we have initiated a genomic approach for the characterization of uncultivated microorganisms from soil. We have developed a procedure based on a two-phase electrophoresis technique that allows the fast and reliable purification of concentrated and clonable, high molecular weight DNA. From this DNA we have constructed complex large-insert genomic libraries. Using archaea-specific ...
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Ynthetic crystals of nickel ammonium sulphate (NiSO4(NH4)2.6H2O) Such crystals are made by precipitation from a solution of nickel in sulphuric acid. In this example the crystal precipitate is realized on a rock subsurface to suggest a natural origin of the crystals. Actual width of sample is 230 mm - Stock Image C026/8100