Dnase

Manufactured by Beyotime

Sourced in China

DNase is a lab equipment product used in molecular biology and biochemistry. It is an enzyme that catalyzes the degradation of DNA molecules by hydrolyzing the phosphodiester bonds between the nucleotide residues.

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Lab products found in correlation

Trizol reagent, by Thermo Fisher Scientific (2 mentions) Nintedanib, by Merck Group (1 mentions) Anti collagen 1, by Affinity Biosciences (1 mentions) Depc treated water, by Thermo Fisher Scientific (1 mentions) Hrp labeled goat anti rabbit igg, by Abcam (1 mentions) Hrp labeled goat anti mouse igg, by Abcam (1 mentions) Anti α actin antibody, by Santa Cruz Biotechnology (1 mentions) Ripa lysis buffer middle, by Beyotime (1 mentions) Anti fibronectin, by Affinity Biosciences (1 mentions) Wnt3a, by Thermo Fisher Scientific (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Fastking gdna dispelling rt supermix, by Tiangen Biotech (1 mentions) Bca kit, by Beyotime (1 mentions) Bleomycin, by Nippon Kayaku (1 mentions) Reverse transcription kit, by Takara Bio (1 mentions) Nanodrop nd 2000 spectrophotometer, by Thermo Fisher Scientific (1 mentions) Trizol, by Takara Bio (1 mentions) Protein a g magnetic beads, by Merck Group (1 mentions) Collagenase, by Merck Group (1 mentions)

Close spelling variants of this product

DNase I Protease K without DNase DNase free RNase Proteinase K without DNase DEPC water (DNase/RNase free) DNase I enzyme DNase-free proteinase K DNase-free protease K RNase-free DNase I

6 protocols using dnase

eDNA Contamination Mitigation in Genetic Studies

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As contamination is a concern in presence–absence genetic studies, in particular eDNA studies, ultrapure water was filtered in the field and tested along with other filters to allow us to identify any equipment or background contamination. All equipment was sterilized using a 5-min exposure to a 10% bleach solution before sampling. Scissors, tweezers, EP tubes, pipettes, tips, and glass cores of vacuum pumps were sterilized at 121°C for 30 min before use. The work area was cleaned using RNase, DNase, RNA, and DNA Away Reagent (Beyotime Biotechnology, Shanghai).
Each eDNA sample was PCR-amplified in triplicate to account for stochasticity in amplifications of low-quality/quantity DNA. A clear gel band in any replicate indicated a positive amplification, which was purified and then subjected to Sanger sequencing. The likely origin of each amplified sequence was determined by a BLAST search in the NCBI non-redundant database, with the top-hit species (based on e-value) representing its origin.

Wang J., Liu P., Chang J., Li C., Xie F, & Jiang J. (2021). Development of an eDNA metabarcoding tool for surveying the world’s largest amphibian. Current Zoology, 68(5), 608-614.

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Nintedanib and Wnt3a Signaling Pathway

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Nintedanib (>99%) was purchased from HWRK Chem Co., Ltd. (Beijing, China). For each experiment, Nintedanib was freshly prepared by dissolving it in DMSO (Sigma-Aldrich, USA). Bleomycin (BLM) was acquired from Nippon Kayaku (Tokyo, Japan). Wnt3a was purchased from Peprotech (Texas, USA). TRIzol reagent and DEPC-treated water were obtained from Thermo Fisher Scientific corporation (Waltham, USA). RNase, DNase, and DNA Away H₂O were purchased from Beyotime Biotechnology (Beijing, China). FastKing gDNA Dispelling RT SuperMix was obtained from Tiangen Biotech Co., Ltd. (Beijing, China). The inhibitor KX2-391, RIPA lysis buffer (middle) and the BCA kit were purchased from Beyotime Biotechnology (Beijing, China). The primary antibodies described in the study included anti-fibronectin, anti-collagen I, and anti-β-tubulin antibodies (Affinity Biosciences, USA); anti-GAPDH, anti-p(Y416)-Src, and anti-Src antibodies (Cell Signalling Technology, USA); anti-α-actin antibody (Santa Cruz Biotechnology, USA); anti-β-catenin and anti-lamin B1 antibodies (ProteinTech Group, China); and anti-p(Y654)-β-catenin antibody (Immunoway Biotechnology, China). The secondary antibodies HRP-labeled goat anti-rabbit IgG and HRP-labeled goat anti-mouse IgG were from Abcam (Cambridge, UK).

Li X., Liu X., Deng R., Gao S., Yu H., Huang K., Jiang Q., Liu R., Li X., Zhang L., Zhou H, & Yang C. (2020). Nintedanib Inhibits Wnt3a-Induced Myofibroblast Activation by Suppressing the Src/β-Catenin Pathway. Frontiers in Pharmacology, 11, 310.

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Isolation of Cytosolic and Nuclear Fractions

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The cytosolic and nuclear fractions from cultured cells were obtained as previously described [28 (link)]. Briefly, cell samples were incubated on ice for 10 min in cytosolic lysis buffer (10 mM Tris-HCl pH 8.0, 60 mM KCl, 1 mM EDTA, protease inhibitor and 0.5% NP-40) and gently triturated using a 26-gauge needle. After centrifugation at 3000 rpm for 5 min at 4 °C, the supernatant was collected as the cytosolic fraction. The pellet was re-suspended in nuclear lysis buffer (20 mM Tris-HCl, pH 8.0, 420 mM NaCl, 0.2 mM EDTA, 25% glycerol, 1.5 mM MgCl₂, protease inhibitor, 1% Triton-X100 in ultrapure water) supplemented with DNase (Beyotime, China), incubated on ice for 30 min, repeatedly triturated, and centrifuged at 14,000 rpm for 10 min at 4 °C. The resulting supernatant was retained as the nuclear fraction.

Tao X., Sun M., Chen M., Ying R., Su W., Zhang J., Xie X., Wei W, & Meng X. (2019). HMGB1-modified mesenchymal stem cells attenuate radiation-induced vascular injury possibly via their high motility and facilitation of endothelial differentiation. Stem Cell Research & Therapy, 10, 92.

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qRT-PCR Analysis of Gene Expression

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The qRT-PCR assay was conducted to analyze the gene abundances following our recent studies [6 (link)]. Total RNA was extracted using Trizol (#9108, TaKaRa, Tokyo, Japan). The integrity of the total RNA was measured using agarose gel electrophoresis (Supplementary Materials Figure S1). The purity of the total RNA was determined using a Nanodrop ND-2000 spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA) by measuring OD₂₆₀, OD₂₈₀, and OD₂₃₀ (OD₂₆₀/₂₈₀ ≥ 1.8, OD₂₆₀/₂₃₀ ≥ 1.5) (Supplementary Materials Table S2), Total RNA was treated with DNase (#D7076, Beyotime Biotechnology, Shanghai, China) and then reverse transcribed into cDNA using a reverse transcription kit (#RR036, TaKaRa). qRT-PCR was conducted in a 25 μL reaction system which contained qPCR Mix (#RR430, TaKaRa), diluted cDNA template, forward and reverse primers, and double distilled H₂O. In addition, no reverse transcriptase (NRT) and no-template control (NTC) analysis was performed. Gene-specific primers are provided in Supplementary Materials Table S3. We normalized the relative mRNA abundance values to the housekeeping genes (β-actin and gapdh), and calculated fold changes using 2^−ΔΔCt method.

Xu Y.C., Zheng H., Guo J.C., Tan X.Y., Zhao T., Song Y.F., Wei X.L, & Luo Z. (2023). Effects of Different Dietary Zinc (Zn) Sources on Growth Performance, Zn Metabolism, and Intestinal Health of Grass Carp. Antioxidants, 12(9), 1664.

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RNA m6A Enrichment and Analysis

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Total RNA was extracted using TRIzol, and DNAse (D7073, Beyotime, China) was added for DNA removal. Subsequently, the MeRIP assay was performed using an m⁶A RNA Enrichment Kit (E1610S, New England Biolabs, Inc., Ipswich, MA, USA). RNA from PTECs was prepared and incubated with protein A/G magnetic beads (16-663X, Sigma-Aldrich, Germany) pre-conjugated with an antibody against m⁶A or IgG. The co-precipitated RNAs were eluted from the beads, purified, and dissolved in RNase-free water. The binding RNA targets were analyzed by PCR (29 (link)).

Li X., Liang Q., Liu L., Chen S., Li Y, & Pu Y. (2024). FTO attenuates TNF-α-induced damage of proximal tubular epithelial cells in acute pancreatitis-induced acute kidney injury via targeting AQP3 in an N6-methyladenosine-dependent manner. Renal Failure, 46(1), 2322037.

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Murine Lung Isolation and Cell Separation

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Murine lung was removed from mice with perfusion with 1× PBS (twice), cut into small pieces, incubated in RPMI 1640 containing 20 µg/mL DNase (Beyotime) and 30 µg/mL collagenase (Sigma) for 40 min at 37°C, and passed through a 200 mesh nylon cell lter. Then the cells were washed with RPMI 1640 containing 10% FBS two times. The erythrocytes were lysed with Ammonium-Chloride-Potassium (ACK) lysis buffer. The isolation of liver monocytes and splenocytes was performed as previously described [27] (link).

Lou Y., Xie D., Huang X., Zheng M., Zhang T., & Xu J. (2021). Vibrio vulnificus Hemolysin Induces TNF-α Independent ROS Production by Primary Murine Macrophages.

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