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Dna quantification kit

Manufactured by Merck Group
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The DNA quantification kit is a laboratory tool designed to accurately measure the concentration of DNA in a sample. The kit provides the necessary reagents and protocols to determine the quantity of DNA present, without making claims about its intended use or applications.

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Lab products found in correlation

Chondroitin sulfate, by Merck Group (2 mentions) Proteinase k, by Roche (2 mentions) Polyvinyl alcohol (pva), by Fujifilm (1 mentions) Recombinant human insulin, by Fujifilm (1 mentions) Cellstain double staining kit, by Dojindo Laboratories (1 mentions) Sodium dihydrogen phosphate nah2po4, by Fujifilm (1 mentions) L cysteine hydrochloride monohydrate, by Merck Group (1 mentions) Ethylenediaminetetraacetic acid edta, by Merck Group (1 mentions) Papain, by Merck Group (1 mentions) Dulbecco s modified eagle s medium dmem, by Merck Group (1 mentions) Micro bca protein assay kit, by Thermo Fisher Scientific (1 mentions) Lysis buffer, by Merck Group (1 mentions) Hoechst 33258 dye, by Merck Group (1 mentions) Varioskan flash multimode reader, by Thermo Fisher Scientific (1 mentions) Hoechst 33258, by Merck Group (1 mentions)

7 protocols using dna quantification kit

1

Quantification of Glycosaminoglycan Content

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Constructs were lyophilized and digested in 1 mL of Proteinase K (Roche, Indianapolis, IN, USA, Cat: 03115828001) working solution (1 : 200 Proteinase K to ultra-pure distilled water and 10 mM Tris-HCl) for 20 h (on a shaker at 250 rpm, 60 °C). GAG content was measured in a 96-well plate by a colorimetric DMMB (Sigma-Aldrich, Cat: 341088), using chondroitin sulfate (Sigma-Aldrich, Cat: c4384) for the standard curve. Plates were read at 530 nm wavelength on an Enspire Plate Reader. GAG content was normalized to DNA content using the Sigma-Aldrich DNA Quantification Kit (Cat: DNAQF).
Tang S., Salazar-Puerta A., Richards J., Khan S., Hoyland J.A., Gallego-Perez D., Walter B., Higuita-Castro N, & Purmessur D. (2021). NON-VIRAL REPROGRAMMING OF HUMAN NUCLEUS PULPOSUS CELLS WITH FOXF1 VIA EXTRACELLULAR VESICLE DELIVERY: AN IN VITRO AND IN VIVO STUDY. European cells & materials, 41, 90-107.
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2

PLGA-Based Insulin Delivery System

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PLGA (copolymer composition ratio of 50 : 50, weight average molecular weight of 20 kDa, and inherent viscosity of 0.187 to 0.229 dL/g), methylene chloride (CH2Cl2), polyvinyl alcohol (86–90 mol% hydrolysis), recombinant human insulin, hydrochloric acid (HCl), sodium hydroxide (NaOH), absolute ethanol (99.5%), N-hydroxysuccinimide esters (NHS), 25% glutaraldehyde solution, and sodium dihydrogen phosphate (NaH2PO4) were obtained from Wako Pure Chemicals Ltd., Japan. L-cysteine hydrochloride monohydrate (minimum 98%), ethylene diamine tetra acetic acid (EDTA), papain, DNA quantification kit, Dulbecco's Modified Eagle's Medium (DMEM), growth supplements, and antibiotics were obtained from Sigma-Aldrich, USA. Phosphate buffer saline (10x, pH = 7.4) was obtained from Nacali Tesque Inc., Japan. Porcine collagen type-1 was obtained from Nitta Gelatin, Japan. 1-Ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride (EDC/EDAC) was obtained from Peptide Institute Inc., Japan. Cellstain Double Staining Kit was obtained from Dojindo Laboratories, Japan. Micro BCA protein assay Kit was obtained from Pierce Biotechnology, USA. All the materials in this study were used as received without further purification. Molecular biology grade milli-Q water from millipore water system (Millipore Corporation, USA) was used for preparation of all the solutions and reagents.
Nanda H.S., Chen S., Zhang Q., Kawazoe N, & Chen G. (2014). Collagen Scaffolds with Controlled Insulin Release and Controlled Pore Structure for Cartilage Tissue Engineering. BioMed Research International, 2014, 623805.
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3

Quantification of Glycosaminoglycan Content

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Constructs were lyophilized and digested in 1 mL of Proteinase K (Roche, Indianapolis, IN, USA, Cat: 03115828001) working solution (1 : 200 Proteinase K to ultra-pure distilled water and 10 mM Tris-HCl) for 20 h (on a shaker at 250 rpm, 60 °C). GAG content was measured in a 96-well plate by a colorimetric DMMB (Sigma-Aldrich, Cat: 341088), using chondroitin sulfate (Sigma-Aldrich, Cat: c4384) for the standard curve. Plates were read at 530 nm wavelength on an Enspire Plate Reader. GAG content was normalized to DNA content using the Sigma-Aldrich DNA Quantification Kit (Cat: DNAQF).
Tang S., Salazar-Puerta A., Richards J., Khan S., Hoyland J.A., Gallego-Perez D., Walter B., Higuita-Castro N, & Purmessur D. (2021). NON-VIRAL REPROGRAMMING OF HUMAN NUCLEUS PULPOSUS CELLS WITH FOXF1 VIA EXTRACELLULAR VESICLE DELIVERY: AN IN VITRO AND IN VIVO STUDY. European cells & materials, 41, 90-107.
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4

Organoid-based GLP-1 secretion assay

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Organoids cultured in 96 well plates for 48 hours were washed 3 times in
Advanced DMEM/F12 with Hepes and L-glutamine. Medium with stimulants (L3740, LCA
or BIMU 8) and control medium (without stimuli) was added to the wells with 5
replicates for each condition (unless otherwise indicated), and slowly shaken on
an orbital shaker after addition of the medium and before the collection. After
2 hours at 37°C, the medium was collected and assayed for GLP-1 content
using GLP-1 V-plex MSD kit (Mesoscale). For glucose stimulation, the organoids
were washed and pre-incubated with DMEM supplemented with 2% FBS containing no
glucose and glutamine for 1 hour. Then DMEM containing 18 mM glucose and 2 mM
L-glutamine or DMEM with no stimulants (control) was added. Experiment was
performed 3-4 times. The values were normalized to the DNA content in the wells
measured with DNA quantification kit (Sigma).
Lund M.L., Sorrentino G., Egerod K.L., Kroone C., Mortensen B., Knop F.K., Reimann F., Gribble F.M., Drucker D.J., de Koning E.J., Schoonjans K., Bäckhed F., Schwartz T.W, & Petersen N. (2020). L-cell differentiation is induced by bile acids through GPBAR1 and paracrine GLP-1 and serotonin signaling. Diabetes, 69(4), 614-623.
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5

Cardiomyocyte Protein-DNA Ratio Analysis

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After standard culture procedures, the ratio of total protein to DNA of cardiomyocytes was analysed. In brief, cells were rinsed in cold PBS twice and scraped with 100 μL of lysis buffer (150 mmol/L NaCl, 15 mmol/L sodium citrate and 0.25% SDS; pH = 7.0). The collected cells were immediately frozen and stored at −20°C. Then samples were thawed and vortexed, and 1 μL sample was applied to determine the total protein content using BCA method. DNA concentrations were detected by using a DNA Quantification Kit (Sigma‐Aldrich).
Zhang M., Jiang Y., Guo X., Zhang B., Wu J., Sun J., Liang H., Shan H., Zhang Y., Liu J., Wang Y., Wang L., Zhang R., Yang B, & Xu C. (2019). Long non‐coding RNA cardiac hypertrophy‐associated regulator governs cardiac hypertrophy via regulating miR‐20b and the downstream PTEN/AKT pathway. Journal of Cellular and Molecular Medicine, 23(11), 7685-7698.
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6

DNA Quantification Protocol Using Hoechst Assay

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A DNA quantification kit (Sigma) was used according to the manufacturer’s protocol. Briefly, the cell-spheres complexes were rinsed with PBS once, homogenized in 1×DNA assay buffer and equal volume of a lysis buffer (Sigma) and then incubated at 37 °C for 1 hr. Cell lysate was centrifuged at 5,000 g at room temperature for 3 min. The supernatant was collected for DNA content determination using fluorescence assay with Hoechst 33258 dye (Sigma) under a Varioskan Flash multimode reader (Thermo Scientific, Wyman Street Waltham, MA ). The excitation wavelength was 360 nm and emission wavelength was 460 nm.
Kuang R., Zhang Z., Jin X., Hu J., Gupte M.J., Ni L, & Ma P.X. (2015). Nanofibrous Spongy Microspheres Enhance Odontogenic Differentiation of Human Dental Pulp Stem Cells. Advanced healthcare materials, 4(13), 1993-2000.
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7

Measuring Osteoblast Proliferation on AZ31 Substrates

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The total cellular DNA content levels of MC3T3E1 osteoblast cells was determined as a measure of cell proliferation [12] (link). Cells were cultured on the coated and uncoated AZ31 substrates at a density of 2.5 × 10 4 cells.cm -2 for 3, 7
Fig. 1 ATR-IR of spectra different substrates and 14 days in a differentiation medium. The differentiation medium was prepared by adding 50 µM ascorbic acid, 100 nM dexamethasone and 10 mM β-glycerophosphate in F12/ DMEM [12] (link). After 3, 7 and 14 days of culture, substrates were washed with PBS and the cells were lysed using cell lysis buffer. The fluorescent dye, Hoechst 33258 (Sigma-Aldrich) was used to quantify the cellular DNA according to the manufacturer's instructions (DNA quantification kit, Sigma-Aldrich).
Agarwal S., Labour M.N., Hoey D., Duffy B., Curtin J, & Jaiswal S. (2018). Enhanced corrosion resistance and cytocompatibility of biomimetic hyaluronic acid functionalised silane coating on AZ31 Mg alloy for orthopaedic applications. Journal of materials science. Materials in medicine, 29(9).
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