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Meiotic double-strand DNA breaks (DSBs), the lesions that initiate meiotic recombination at the HIS4 recombination hot spot, occur in a region upstream of the coding sequence associated with multiple DNase I-hypersensitive sites. Mutations in transcription factors that lead to loss of the DSBs result in the loss of some but not all DNase I-hypersensitive sites in the upstream region. A meiosis-specific change in chromatin structure is detected in strains with the wild-type hot spot but not in strains with alterations that elevate or reduce hot spot activity. The position and intensity of micrococcal nuclease-hypersensitive sites correlate poorly with the sites of DSB formation. ...
DNaseI hypersensitivity - posted in DNA Methylation and Epigenetics: Hi, Does anybody know if you can look at DNaseI hypersensitivity in frozen PBMCs or do the PBMCs need to be from fresh blood. Many thanks.
Alzheimers disease is the most common cause of dementia and is characterized by a progressive loss of brain tissue leading to amyloid-β accumulation and severe decline in cognitive function. The cause of Alzheimers disease is poorly understood, and available treatments are limited in their efficacy, particularly for patients with more severe symptoms. We report the case of a 77-year-old Caucasian man with severe dementia and behavioral disturbance secondary to Alzheimers disease treated with memantine who began adjunct treatment with deoxyribonuclease I. Prior to initiation of deoxyribonuclease I treatment, our patient appeared to be in a stuporous state, with a Mini-Mental State Examination score of 3 and a Functional Assessment Staging Test score of 7. After obtaining informed consent from family members, we started administration of 120 mg of deoxyribonuclease I per day (1500 KU/mg) for treatment of severe cognitive impairment. Our patient began to demonstrate rapid, considerable improvement in
DNase I hypersensitive site assay is perhaps the most widely used assay for detection of changes in chromatin structure. Studies of many spatially or temporally regulated genes, such as P-globin, have revealed the correlation of formation of DNase I hypersensitive sites with the status of transcription (22). This assay has also been used extensively in analyses of chromatin structure in injected Xenopus oocytes (24). Although the mechanism for the formation of DNase I hypersensitive sites is still not fully understood, the detection of DNase I hypersensitive sites are widely interpreted as results of chromatin remodeling induced by the binding of transcription factors.. 1. Inject 15-20 oocytes with DNA and with or without mRNA encoding TR/RXR (100 ng/ pL, 27.6 nL/oocyte) and treated with or without 50 nM T3. Incubate the oocytes overnight at 18°C incubator.. 2. Collect healthy oocytes and wash the oocytes once with 500 pL of MBSH buffer.. 3. To 15-20 oocytes, add 240 pL of DNase I buffer. ...
We have shown previously that fatty acid synthase (FAS) gene expression is positively regulated by glucose in rat adipose tissue and liver. In the present study, we have identified in the first intron of the gene a sequence closely related to known glucose-responsive elements such as in the L-pyruvate kinase and S14 genes, including a putative upstream stimulatory factor/major late transcription factor (USF/MLTF) binding site (E-box) (+ 292 nt to + 297 nt). Location of this sequence corresponds to a site of hypersensitivity to DNase I which is present in the liver but not in the spleen. Moreover, using this information from a preliminary report of the present work, others have shown that a + 283 nt to + 303 nt sequence of the FAS gene can confer glucose responsiveness to a heterologous promoter. The protein binding to this region has been investigated in vitro by a combination of DNase I footprinting and gel-retardation experiments with synthetic oligonucleotides and known nuclear proteins. ...
Recent genome-wide analyses of yeast and human chromatin revealed the widespread prevalence of DNase I hypersensitive sites (DNase I HSs) at gene regulatory regions with possible roles in eukaryotic gene regulation. The presence of DNase I HSs in plants has been described for only a few genes, and we analyzed the chromatin structure of an 80 kb genomic region containing 30 variably expressed genes by DNase I sensitivity assay at 500 bp resolution in Arabidopsis. Distinct DNase I HSs were found at the 5′ and/or 3′ ends of most genes irrespective of their expression levels. Further analysis of well-characterized genes showed that the DNase I HSs occurred near cis-regulatory elements in the promoters of these genes. Upon transcriptional activation of a heat-inducible gene, the DNase I HS was extended into the vicinity of a cis-element and adjacent TATA element in the promoter. Concomitant with this change in DNase I HS, histones were acetylated, removed from the promoter, and a transcription ...
Although it has been assumed that the deregulated bcl-2 expression in t(14;18) cells is mediated in part by the immunoglobulin heavy chain gene regulatory region, this has not been demonstrated, nor was it known which elements of that region were responsible for the deregulation. In these studies, we found that the four DNase I-hypersensitive regions within the IgH 3′ enhancer were able to activate the bcl-2 promoter in the t(14;18) cell line DHL-4. Of those four hypersensitive regions, we demonstrated that HS4 had the most influence on bcl-2 promoter activity. This is similar to the situation in pre-B and plasmacytoma cells, where HS4 is the most active enhancer region. We also showed that the HS1,2 region was capable of activating the promoter independently. By itself, HS3 increased bcl-2 promoter activity by only a minor amount. Other studies of HS3 have shown that it is contains elements that act as negative effectors of the IgH 3′ enhancer (37) , and preliminary studies in our ...
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Deoxyribonuclease I, Recombinant, AOF - Instruments Consumables Reagents Advanced BioMatrix,RANDOX,RANDOX ELISA,Biomedical, biochemical reagents, laboratory supplies, equipment, antibodies, ELISA kits, diagnostic reagents, methods of experimental techniques, general analytical instruments, material testing instruments and equipment, used laboratory equipment, instruments and equipment, life sciences, environmental monitoring equipment , measurement, measuring instruments, rotating wall bioreactor, three-dimensional tissue / stem cell culture system; microcapsule
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... - Instruments Consumables Reagents Advanced BioMatrix,RANDOX,RANDOX ELISA,Biomedical, biochemical reagents, laboratory supplies, equipment, antibodies, ELISA kits, diagnostic reagents, methods of experimental techniques, general analytical instruments, material testing instruments and equipment, used laboratory equipment, instruments and equipment, life sciences, environmental monitoring equipment , measurement, measuring instruments, rotating wall bioreactor, three-dimensional tissue / stem cell culture system; microcapsule
We analyzed genome-wide association studies (GWASs), including data from 71,638 individuals from four ancestries, for estimated glomerular filtration rate (eGFR), a measure of kidney function used to define chronic kidney disease (CKD). We identified 20 loci attaining genome-wide-significant evidence of association (p , 5 × 10(-8)) with kidney function and highlighted that allelic effects on eGFR at lead SNPs are homogeneous across ancestries. We leveraged differences in the pattern of linkage disequilibrium between diverse populations to fine-map the 20 loci through construction of "credible sets" of variants driving eGFR association signals. Credible variants at the 20 eGFR loci were enriched for DNase I hypersensitivity sites (DHSs) in human kidney cells. DHS credible variants were expression quantitative trait loci for NFATC1 and RGS14 (at the SLC34A1 locus) in multiple tissues. Loss-of-function mutations in ancestral orthologs of both genes in Drosophila melanogaster were associated with ...
Monocyte differentiation into macrophages represents a cornerstone process for host defense. Concomitantly, immunological imprinting of either tolerance or trained immunity determines the functional fate of macrophages and susceptibility to secondary infections. Transcriptomes (RNA-Seq) and epigenomes (ChIP-Seq H3K4me1,H3K4me3,H3K27ac) in four primary cell types: monocytes, in vitro differentiated naive, tolerized and trained macrophages were characterized. Inflammatory and metabolic pathways were modulated in macrophages, including decreased inflammasome activation, and pathways functionally implicated in trained immunity were identified. Strikingly, B-glucan training elicits an exclusive epigenetic signature, revealing a complex network of enhancers and promoters. Analysis of transcription factor motifs in DNase I hypersensitive sites at cell-type specific epigenetic loci unveiled differentiation and treatment specific repertoires. Altogether, this study provides a resource to understand the ...
The transcription factor RUNX1/AML1 is an important regulator of hematopoiesis and RUNX1 gene is one of the most frequent target of chromosomal translocations in cells of the myeloid lineage [1]. Interestingly, the RUNX1 gene covers 260 kbp of chromosome 21 but surprisingly, all genomic breakpoints for the leukemia causing translocations (8; 21) and (16;21) are found in intron 5 of the gene [2]. Presently, factors involved in maintaining the structural integrity and/or enhancing susceptibility of these regions to undergo recombination are unknown. Moreover, the breakpoint junctions are devoid of common DNA motifs that can explain the high recombination frequency observed. Interestingly however, topoisomerase II and DNase I hypersensitive sites have been found to correlate with breakpoints suggesting that chromatin organization may be responsible for, or contribute to, chromosomal translocation formation [3, 4]. DNA regions that exhibit DNase I hypersensitivity have been extensively associated ...
InterPro provides functional analysis of proteins by classifying them into families and predicting domains and important sites. We combine protein signatures from a number of member databases into a single searchable resource, capitalising on their individual strengths to produce a powerful integrated database and diagnostic tool.
We categorized human genome 100kb non-overlapping segments by their DNAse Hypersensitive Sites (DHS) counts based on data in (Sheffield et al., 2011). They fit a Weibull long tail distribution with a peak at around 14 DHSs per bin. The few (around 50) bin
DNase I (RNase-free) (100 U/ l) /10X DNase I Buffer enzyme ZE1007 DNase I (RNase-free) (100 U/ l) /10X DNase I Buffer enzyme ZE1007
Recombinant human deoxyribonuclease I (rhDNase) is the mucolytic agent most widely used for the treatment of respiratory disease in cystic fibrosis. However, rhDNase is rapidly cleared from the lungs which implies a high dosing frequency and limited patient adherence. The aim of this study was to produce a long-acting PEGylated derivative of rhDNase presenting a preserved enzymatic activity. Site-specific PEGylation on the N-terminal (N-ter) leucine residue of rhDNase was achieved by reductive alkylation at acidic pH using linear 20kDa, linear 30kDa or two-arm 40kDa polyethylene glycol (PEG) propionaldehydes. Yields of mono-PEGylated products ranged between 45% and 61%. Conjugation to PEG fully preserved the secondary structure and the in vitro enzymatic activity of the native protein. These properties offer interesting perspectives for in vivo inhalation studies of the PEGylated enzyme. ...
951 1234 DNase hypersensitive site 4 4550 4775 DNase hypersensitive site 3 8486 8860 DNase hypersensitive site 2 12752 13769 DNase hypersensitive site 1 116 431 Right Alu 1968 2258 Left Alu 5605 5918 Right Alu 8019 8314 Right Alu 10612 10924 Right Alu 12912 13066 Left L1 14836 15071 Right L1 16918 17218 Left Alu 17940 18231 Right Alu 19486 21080 epsilon-globin gene 19486 19632 Exon 1 19755 19977 Exon 2 20833 21080 Exon 3 23118 31136 Left L1 25885 27987 Left InnerL1 32407 32711 Right Alu 32986 33101 Left L1 34478 36069 G-gamma-globin gene 34478 34622 Exon 1 34745 34967 Exon 2 35854 36069 Exon 3 37921 38039 Left L1 39414 40985 A-gamma-globin gene 39414 39558 Exon 1 39681 39903 Exon 2 40770 40985 Exon 3 42695 43274 Left L1 44788 45108 Right Alu 45658 47272 eta-globin pseudo gene 45658 45800 Exon 1 45922 46144 Exon 2 46997 47272 Exon 3 50895 51198 Left Alu 51976 52276 Right Alu 53222 53540 Left L1 54740 56389 delta-globin gene 54740 54881 Exon 1 55010 55232 Exon 2 56131 56389 Exon 3 62137 63742 ...
He, Housheng Hansen, Clifford A Meyer, Henry Long, X Shirley Liu, and Myles Brown. 2013. A closer look into dnase i hypersensitivity. Epigenetics & Chromatin 6(Suppl 1): P25. ...
TY - JOUR. T1 - Interleukin-8 gene expression in human bronchial epithelial cells*. AU - Nakamura, H.. AU - Yoshimura, K.. AU - Jaffe, H. A.. AU - Crystal, Ronald. PY - 1991/1/1. Y1 - 1991/1/1. N2 - The capacity of cells of the human bronchial epithelium to express the gene for interleukin-8 (IL-8) was evaluated in bronchial epithelium derived cell lines, HS-24 and BET-1A, using tumor necrosis factor-α (TNF) as a model inflammatory stimulus. As in other epithelium, TNF markedly increased the level of the 1.8-kilobase IL-8 mRNA transcripts in both bronchial epithelial cell lines. In HS-24 cells, nuclear run-on analyses showed the IL-8 gene transcription rate was dramatically increased, more than 30-fold, after TNF stimulation. The half-life of IL-8 mRNA transcripts in these cells was approximately 40 min and did not change after TNF stimulation, suggesting that TNF up-regulated IL-8 gene expression mainly at the transcriptional level. DNase I hypersensitivity site mapping of chromatin DNA in ...
Four DNaseI hypersensitive (HS) chromatin regions were found in the uteroglobin locus located at -3.7, -2.4, -0.1 and +4.1 kb with respect to the transcription start site of the gene. The three sites upstream of the gene are only detected in the hormonally stimulated endometrium and disappear after hormone withdrawal, whereas the site at +4.1 is also found in tissues that do not express uteroglobin. In the -2.4 HS region, which is strictly dependent on progesterone treatment, three DNaseI sites are clustered within a 240 bp DNA segment that contains 20 imperfect repeats of an octanucleotide motif. Upstream of the uteroglobin gene there are three regions containing binding sites for the glucocorticoid and the progesterone receptors, located at -3.7, -2.6/-2.7 and -2.4. The -2.4 region contains two binding sites for the hormone receptors flanking the central HS site. In footprinting experiments with naked DNA binding of the receptor also renders this site more susceptible towards digestion with DNaseI.
Profacgen provides professional DNase I footprinting assay service for the identification of exact binding sites of DNA-binding proteins.
The binding of MC1 protein, the major chromosomal protein of the archaebacterium Methanosarcina sp. CHTI 55, to the region preceding the strongly expressed genes encoding methyl coenzyme reductase in a closely related micro-organism has been investigated. By gel retardation and DNAase I footprinting assays, we identified a preferential binding sequence in an open reading frame of unknown function. The large area of DNA protected against DNAase I is interrupted by a strong cleavage enhancement site on each strand. By circular permutation assays, we showed that the DNA bends upon MC1 binding. Furthermore we observed that the presence of a sequence outside the binding site can induce an unusual electrophoretic behaviour in some complexes. ...
The dinuclear iron(II) supramolecular helicates [Fe2L3]Cl-4 (L= C25H20N4) bind to DNA through noncovalent (i. e., hydrogen-bonding, electrostatic) interactions and exhibit antimicrobial and anticancer effects. In this study, we show that the helicates condense plasmid DNA with a much higher potency than conventional DNA-condensing agents. Notably, molecules of DNA in the presence of the M enantiomer of [Fe2L]Cl-4 do not form intermolecular aggregates typically formed by other condensing agents, such as spermidine or spermine. The helicates inhibit the activity of several DNA-processing enzymes, such as RNA polymerase, DNA topoisomerase I, deoxyribonuclease I, and site-specific restriction endonucleases. However, the results also indicate that the DNA condensation induced by the helicates does not play a crucial role in these inhibition reactions. The mechanisms for the inhibitory effects of [Fe2L3]Cl-4 helicates on DNA-related enzymatic activities have been proposed ...
Thermo Scientific DNase I, RNase-free is an endonuclease that digests single- and double-stranded DNA. It hydrolyzes phosphodiester bonds producing mono- and oligodeoxyribonucleotides with 5-phosphate and 3-OH groups.. The enzyme activity is strictly dependent on Ca2+ and is activated by Mg2+ or Mn2+ ions.. ...
View mouse Dnase1l2 Chr17:24440081-24443105 with: phenotypes, sequences, polymorphisms, proteins, references, function, expression
TY - JOUR. T1 - Clinical use of dornase alfa is associated with a slower rate of FEV 1 decline in cystic fibrosis. AU - Konstan, Michael W.. AU - Wagener, Jeffrey S.. AU - Pasta, David J.. AU - Millar, Stefanie J.. AU - Jacobs, Joan R.. AU - Yegin, Ashley. AU - Morgan, Wayne J. PY - 2011/6. Y1 - 2011/6. N2 - Objectives Randomized controlled trials of dornase alfa have shown forced expiratory volume in 1 sec (FEV1) to improve in patients with cystic fibrosis (CF) but have not assessed change in the rate of lung function decline. We assessed the relationship of dornase alfa use and FEV1 decline using the Epidemiologic Study of Cystic Fibrosis (ESCF). Methodology Patients aged 8-38 years who had been enrolled in ESCF for 2 years when initially treated with dornase alfa were selected if they remained on treatment during the following 2 years. A comparator group included patients aged 8-38 who were not yet reported to have received dornase alfa. For each patient we estimated the annual rate of ...
In the era of genome-wide association studies (GWAS) and personalized medicine, predicting the impact of single nucleotide polymorphisms (SNPs) in regulatory elements is an important goal. Current approaches to determine the potential of regulatory SNPs depend on inadequate knowledge of cell-specific DNA binding motifs. Here, we present Sasquatch, a new computational approach that uses DNase footprint data to estimate and visualize the effects of noncoding variants on transcription factor binding. Sasquatch performs a comprehensive k-mer-based analysis of DNase footprints to determine any k-mers potential for protein binding in a specific cell type and how this may be changed by sequence variants. Therefore, Sasquatch uses an unbiased approach, independent of known transcription factor binding sites and motifs. Sasquatch only requires a single DNase-seq data set per cell type, from any genotype, and produces consistent predictions from data generated by different experimental procedures and at
Mapping of DNase I hypersensitive sites (DHSs) is a powerful tool to experimentally identify cis-regulatory elements (CREs). Among CREs, enhancers are abundant and predominantly act in driving cell-specific gene expression. Krüppel-like factors (KLFs) are a family of eukaryotic transcription factors. Several KLFs have been demonstrated to play important roles in hematopoiesis. However, transcriptional regulation of KLFs via CREs, particularly enhancers, in erythroid cells has been poorly understood. In this study, 23 erythroid-specific or putative erythroid-specific DHSs were identified by DNase-seq in the genomic regions of 17 human KLFs, and their enhancer activities were evaluated using dual-luciferase reporter (DLR) assay. Of the 23 erythroid-specific DHSs, the enhancer activities of 15 DHSs were comparable to that of the classical enhancer HS2 in driving minimal promoter (minP). Fifteen DHSs, some overlapping those that increased minP activities, acted as enhancers when driving the corresponding
In vivo studies of gene activation have revealed that changes in chromatin are often associated with transcriptional activation (reviewed in references 21,30, and 44). For example, in erythroid cells where each globin gene is sequentially expressed during development, the human β-globin locus is hypersensitive to nucleases (19, 22). In contrast, the entire β-globin locus is condensed into a nuclease-resistant chromatin structure in nonerythroid tissues, where the genes are not transcribed. Thus, transcriptionally active chromatin domains differ from the bulk of the genome in their susceptibility to digestion by nucleases. Detailed studies of nuclease-hypersensitive sites have suggested that interactions of sequence-specific DNA binding factors with chromatin alter the canonical nucleosomal structure (for example, see references2, 7, and 59). These conformational changes generate transcriptionally competent chromatin templates that are accessible to general transcription factors and the RNA ...
The human growth hormone (hGH) gene cluster includes 5 genes: the pituitary hGH-N gene, and hCS-A, hCS-L, hGH-V, and hCS-B expressed in placenta. These 5 genes are regulated by a locus control region (LCR) located in the far 5 region of this multigene locus. The B-cell specific CD79b/Igß gene is interposed between the hGH LCR and the hGH-N gene. The LCR is made up of 5 DNaseI hypersensitive sites (HS) located between -14.5 kb (HSI) and -32 kb (HSV) 5 to hGH-N. These HS are only present in placenta and/or pituitary. When this 32 kb domain plus the linked GH genes are present in a transgenic construct, transgenic mice reproducibly and robustly express the hGH-N gene specifically in pituitary somatotropes and the hCS genes specifically in the placenta. Expression in both tissues is copy-number-dependent and site-of-integration independent. This expression is paralleled by the establishment of a pituitary-specific 32 kb acetylated chromatin domain encompassing the hGH LCR and the contiguous hGH-N ...
Study Lecture 16 - Type I hypersensitivity - ALLERGY flashcards from M G's class online, or in Brainscape's iPhone or Android app. ✓ Learn faster with spaced repetition.
Treatments for Type I Hypersensitivity including drugs, prescription medications, alternative treatments, surgery, and lifestyle changes.
These tracks complement each other and together can shed much light on regulatory DNA. The histone marks are informative at a high level, but they have a resolution of just ~200 bases and do not provide much in the way of functional detail. The DNase hypersensitivity assay is higher in resolution at the DNA level and can be done on a large number of cell types since its just a single assay. At the functional level, DNase hypersensitivity suggests that a region is very likely to be regulatory in nature, but provides little information beyond that. The transcription factor ChIP assay has a high resolution at the DNA level and, due to the very specific nature of the transcription factors, is often informative with respect to functional detail. However, since each transcription factor must be assayed separately, the information is only available for a limited number of transcription factors on a limited number of cell lines. Though each assay has its strengths and weaknesses, the fact that all of ...
Methodology/Principal Findings: Total nuclease activity and Dnase1 expression and activity was evaluated using in situ and in vitro analyses of kidneys and sera from (NZBxNZW)F1 mice of different ages, and from age-matched healthy controls. Immunofluorescence staining for Dnase1 was performed on kidney biopsies from (NZBxNZW)F1 mice as well as from human SLE patients and controls. Reduced serum Dnase1 activity was observed in both mesangial and end-stage lupus nephritis. A selective reduction in renal Dnase1 activity was seen in mice with massive deposition of chromatin-containing immune complexes in glomerular capillary walls. Mice with mild mesangial nephritis showed normal renal Dnase1 activity. Similar differences were seen when comparing human kidneys with severe and mild lupus nephritis. Dnase1 was diffusely expressed within the kidney in normal and mildly affected kidneys, whereas upon progression towards end-stage renal disease, Dnase1 was down-regulated in all renal compartments. This ...
Find information about cystic fibrosis and Pulmozyme®, including your CF treatments and options for financial support. Indication and Usage Pulmozyme® (dornase alfa) is indicated for daily administration along with standard therapies for the management of cystic fibrosis (CF) patients to improve pulmonary function. In CF patients with an FVC ≥ 40% of predicted, daily administration of Pulmozyme® has also been shown to reduce the risk of respiratory tract infections requiring injectable antibiotics. Important Safety Information Pulmozyme® should not be used in patients who are allergic to any of its ingredients. Patients may experience the following when using Pulmozyme®: change in or loss of their voice, discomfort in the throat, rash, chest pain, red watery eyes, runny nose, lowering of lung function, fever, indigestion, and shortness of breath. There have been no reports of severe allergic reactions caused by the administration of Pulmozyme. Mild to moderate hives and mild skin rash have
Learn what cystic fibrosis is, its signs, symptoms, and possible causes as well as why your doctor may prescribe Pulmozyme® as a treatment. Indication and Usage Pulmozyme® (dornase alfa) is indicated for daily administration along with standard therapies for the management of cystic fibrosis (CF) patients to improve pulmonary function. In CF patients with an FVC ≥ 40% of predicted, daily administration of Pulmozyme® has also been shown to reduce the risk of respiratory tract infections requiring injectable antibiotics. Important Safety Information Pulmozyme® should not be used in patients who are allergic to any of its ingredients. Patients may experience the following when using Pulmozyme®: change in or loss of their voice, discomfort in the throat, rash, chest pain, red watery eyes, runny nose, lowering of lung function, fever, indigestion, and shortness of breath. There have been no reports of severe allergic reactions caused by the administration of Pulmozyme. Mild to moderate hives and
Overview. DNase I from bovine pancreas is a glycoprotein of Mr 37000. A special procedure is used to remove RNases from the DNase preparation. DNase I is a DNA-specific endonuclease that hydrolyzes ds or ssDNA to a mixture of oligo and mononucleotides. The enzyme requires divalent cations for maximal activity. The specificity of the reaction depends on the nature of the cations. In the presence of Mg2+ DNase I cause nicks in dsDNA, while in the presence of Mn2+ the enzyme produces double-stranded breaks.. Suggested Method for the Removal of DNA for RT-PCR. 10 x reaction buffer: 200 mM Tris-HCl, pH 8.3 500 mM KCl 10 mM MnCl2 recommended to add 20 U Protector RNase Inhibitor ...
File Title: Active Chromatin Structure of Saccharomycea cerevisiae: High Mobility Group Proteins, Histone Modifications and DNase I Sensitivity ...
... provided by the Genemart Bioscience Inc.,The Product Intro:DNase I, Bovine Pancreas, >2000U/MG
In article ,6lhd6.14135$KP3.4431209 at news3.rdc1.on.home.com,, the eminent EO at Excite at Home - The Leader in Broadband http://home.com/faster wrote ,Can you be more specific as to how much buffer and suspect to use? How ,about 50 mL buffer with 0.1 gram reagent.? That is a little OTT! 1ug of a Hae III digest of pBR322 (or similar) should be trashed with less than a fraction of a unit of DNAse. As pure DNase is probably 3000u/mg then I would hazard a guess that ng quantities of DNase would be picked up, maybe even pg. Easy to test. Prepare a reaction mix with 1ug of digested plasmid/50ul and dispense 45ul into 10 microtubes. Prepare a 2u/ul stock of DNase in same buffer. Add 5ul (10u) DNase to tube 1. Mix. Remove 5ul with a fresh tip to tube 2. Mix and repeat down the line remembering not to add DNAse to last tube 1st tube has 0.2u DNase/ul 2nd tube 0.02 3rd 0.002 etc etc Incubate 37C for one hour. Run on acrylamide gel, stain with EtBr and see what happens. For increased sensitivity 37C ...
ZR-96 Quick-RNA™ Kit (4 x 96 Preps) [[Includes E1009 x 8: DNase I Set (250 U) w/ 10X Reaction Buffer (1 ml)] Quick-RNA™ MiniPrep Kit (50 Preps) w/Zymo-Spin™ IIICG Columns (Capped) & Spin-Away™ Filters [Includes E1009 x 1: DNase I Se Sample: Quick-RNA™ MiniPrep Kit (10 Preps) Quick-RNA™ MiniPrep Kit (200 Preps) w/Zymo-Spin™ IIICG Columns (Capped) & Spin-Away™ Filters [Includes E1009 x 4: DNase I Set (250 U) w/ 10X Reaction Buffer (1 ml)] ZR small-RNA™ PAGE Recovery Kit (20 Preps) ZR-96 RNA Clean & Concentrator™ Kit (2 x 96 Preps) RNA Shield Purification Kit (50 prep) supplied w/ 50 ml RNA Shield RNA Shield Purification Kit (50 prep) Without RNA Shield ZR Fungal/Bacterial RNA MicroPrep™ Kit (50 Preps) ZR Fungal/Bacterial RNA MiniPrep™ Kit (50 Preps) ZR Tissue & Insect RNA MicroPrep™ Kit (50 Preps) ZR Soil/Fecal RNA MicroPrep™ Kit (50 Preps) Direct-zol™ RNA MiniPrep (50 Preps) w/ Zymo-Spin™ IIC Columns (Capped) [Includes E1009 x 1: DNase I Set (250 U) w/ 10X Reaction ...
Deoxyribonuclease BamHI information including symptoms, causes, diseases, symptoms, treatments, and other medical and health issues.
Define deoxyribonuclease. deoxyribonuclease synonyms, deoxyribonuclease pronunciation, deoxyribonuclease translation, English dictionary definition of deoxyribonuclease. n. DNase. n the full name for DNAase n. deoxyribonuclease: any of several enzymes that break down the DNA molecule into its component nucleotides
The insulating properties required to delimit higher-order chromosomal domains have been shown to be shared by a variety of chromatin boundary elements (BEs). Boundary elements have been described in several species, from yeast to human, and we have previously reported the existence of a class of chromatin BEs in Drosophila melanogaster whose insulating activity requires the DNA-binding protein BEAF (boundary element-associated factor). Here we focus on the characterization of a moderately repeated 1.2 kb DNA sequence that encompasses boundary element 28 (BE28). We show that it directionally blocks enhancer/promoter communication in transgenic flies. This sequence contains a BEAF-binding sequence juxtaposed to an AT-rich sequence that harbors a strong nuclease-hypersensitive site. Using a combination of DNA-protein and protein blotting techniques, we found that this region is recognized by the A+T-binding D1 non-histone chromosomal protein of D. melanogaster, and we provide evidence that D1 and ...
Deoxyribonuclease II (EC 3.1.22.1, DNase II, pancreatic DNase II, deoxyribonucleate 3-nucleotidohydrolase, pancreatic DNase II, acid deoxyribonuclease, acid DNase) hydrolyzes deoxyribonucleotide linkages in native and denatured DNA, yielding products with 3-phosphates. As the name implies, it is more effective at acid pH.. There are several known DNases II, including:. ...
Nucleosomes are important for gene regulation because their arrangement on the genome can control which proteins bind to DNA. Currently, few human nucleosomes are thought to be consistently positioned across cells; however, this has been difficult to assess due to the limited resolution of existing data. We performed paired-end sequencing of micrococcal nuclease-digested chromatin (MNase-seq) from seven lymphoblastoid cell lines and mapped over 3.6 billion MNase-seq fragments to the human genome to create the highest-resolution map of nucleosome occupancy to date in a human cell type. In contrast to previous results, we find that most nucleosomes have more consistent positioning than expected by chance and a substantial fraction (8.7%) of nucleosomes have moderate to strong positioning. In aggregate, nucleosome sequences have 10 bp periodic patterns in dinucleotide frequency and DNase I sensitivity; and, across cells, nucleosomes frequently have translational offsets that are multiples of 10 bp. ...