Nc sirna

Manufactured by Sangon

Sourced in China

NC siRNA is a laboratory reagent used for experimental purposes. It is a non-targeting small interfering RNA (siRNA) designed to serve as a negative control in RNA interference (RNAi) experiments.

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Lab products found in correlation

23 protocols using nc sirna

Modulation of miR-93-5p and PDCD4 in AOH

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A total of 60 adult male SD rats weighing 200–250 g were divided into six groups using a random number table: the control group, AOH group, miR-93-5p agomir group with AOH (miR-93-5p+AOH), miR-93-5p agomir negative control group with AOH (miR-93-5p NC+AOH), siPDCD4 group with AOH (siPDCD4+AOH), and siRNA negative control group with AOH (siRNA NC+AOH).
miR-93-5p agomir (chemically modified miRNA agonist), siPDCD4 (chemically modified small interfering RNA sequence inhibiting PDCD4 expression), agomir NC, and siRNA NC (random sequence) were synthesized by Sangon Biotech. miR-93-5p agomir, siPDCD4, agomir NC, and siRNA NC (0.5 nmol/μl) were injected into the vitreous cavity of the corresponding group of AOH rats 30 min before modeling. Intravitreal injection was performed according to previous literature reports (36 (link), 54 (link)). Briefly, intravitreal injection was performed between two vortex veins about 1 mm from the corneal limbus.

Tan C., Shi W., Zhang Y., Liu C., Hu T., Chen D, & Huang J. (2023). MiR-93-5p inhibits retinal neurons apoptosis by regulating PDCD4 in acute ocular hypertension model. Life Science Alliance, 6(9), e202201732.

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Resveratrol-mediated cellular apoptosis

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Resveratrol (RES, purity >99%), dimethyl sulfoxide (DMSO), annexinV-FITC apoptosis assay kit, superoxide dismutase (SOD) kit, glutathione (GSSH) kit, and ELISA kit for IL-6 detection were purchased from Solarbio (China). GSH and GSSH assay kits were purchased from Beyotime Biotechnology Co., Ltd. Dulbecco's Modified Eagle's Medium (DMEM), fetal bovine serum (FBS), penicillin-streptomycin solution (SP), phosphate-buffered saline (PBS), and trypsin-EDTA (0.25%) were purchased from Gibco (USA). 3-(4,5dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was procured from GEN-view. TRIzol was acquired from Takara Bio (Japan). The BCA protein assay kit was provided by Thermo Fisher (China). siRNA/miRNA in vitro transfection reagents were provided by Yeasen Biotechnology Co., Ltd. (Shanghai). Lastly, siRNA VEGF-2645 and siRNA NC were procured from Sangon Biotech Co., Ltd. (Shanghai).
SH4 ultraviolet light therapy instrument with wavelength ranging between 311 and 313 nm, and having a peak wavelength of 313 nm, was purchased from Shanghai Sigma High-Tech Co., Ltd., and Corneometer CM825 skin moisture content tester was procured from CK (Germany).

Cui B., Wang Y., Jin J., Yang Z., Guo R., Li X., Yang L, & Li Z. (2022). Resveratrol Treats UVB-Induced Photoaging by Anti-MMP Expression, through Anti-Inflammatory, Antioxidant, and Antiapoptotic Properties, and Treats Photoaging by Upregulating VEGF-B Expression. Oxidative Medicine and Cellular Longevity, 2022, 6037303.

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Transfection of miRNA mimics and inhibitors

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Negative control (NC) mimic, miR-486-5p mimic, NC inhibitor, miR-486-5p inhibitor, siRNA-NC and siRNA-RAP1 were synthesized by Sangon Biotech (Shanghai, China). Then they were transfected into indicated cells by using Lipofectamine 2000 Transfection Reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instruction. 48 h after transfection, the transfection efficiency was tested by qRT-PCR.

Ling Q., Wu S., Liao X., Liu C, & Chen Y. (2022). Anesthetic propofol enhances cisplatin-sensitivity of non-small cell lung cancer cells through N6-methyladenosine-dependently regulating the miR-486-5p/RAP1-NF-κB axis. BMC Cancer, 22, 765.

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siRNA Targeting RSF-1 in Cervical Cancer Cells

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siRNA targeting RSF-1 (siRNA-RSF-1) and siRNA negative control (siRNA-NC) were obtained from GenePharma Co. Ltd. (Shanghai, China). The sequences of siRNA-RSF-1 were: sense, 5ʹ-GGAGAAGAAGGGCUAUCUUTT-3ʹ; antisense, 5ʹ-AAGAUAGCCCUUCUUCUCCTT-3ʹ. The sequences of siRNA-NC were: sense, 5ʹ-UUCUCCGAACGUGUCACGUTT-3ʹ; antisense, 5ʹ- ACGUGACACGUUCGGAGAATT-3ʹ. pcDNA3.1 containing RSF-1 plasmid (pcDNA3.1-RSF-1) and the empty vector were purchased from Shanghai Sangon Biotech Co., Ltd (Shanghai, China). siRNA or plasmids were transfected into HeLa and SiHa cells using a Lipofectamine 3000 transfection reagent kit (Invitrogen) according to the manufacturer’s protocol.

Tian J., Kong E., Wang X., Xie Z., Chang C.Y., Sheu J.J., Hao Q, & Sun L. (2020). RSF-1 siRNA Enhances Tumor Radiosensitivity in Cervical Cancer via Enhanced DNA Damage, Cell Cycle Redistribution, and Promotion of Apoptosis. OncoTargets and therapy, 13, 3061-3071.

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Chicken HELZ2 and IFI6 siRNA Knockdown

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Chicken zinc-fingered helicase 2 (HELZ2) siRNA (+): 5’-GCUGUUCCUUGAGGAUUATT-3’, siRNA (-): 5’-UAAUCCUCAAGGGAACAGCTT -3’, chicken interferon-α-inducible protein 6 (IFI6) siRNA (+): 5’-CCACAAA GCCGGUUUCACUTT-3’, siRNA (-): 5’ -AGUGAAACCGGCUUUGUGGTT-3’ and NC siRNA (+): 5’-UUCUCCGAACGUGUCACGUTT-3’, siRNA (-): 5’-ACG UGAC ACGUUCGGAGAATT were purchased from Sangon Biotech (Shanghai, China). Transfection of siRNA was performed using TransIntro™ EL Transfection Reagent(TransGen Biotech, Beijing, China) according to the manufacturer’s instructions. Five hours after transfection, cells were infected with DTMUV at an MOI of 1 or mock-treated, and incubated until harvest at 24, 36 and 48 hpi, respectively.

Xiang C., Yang Z., Xiong T., Wang T., Yang J., Huang M., Liu D, & Chen R. (2022). Construction and Transcriptomic Study of Chicken IFNAR1-Knockout Cell Line Reveals the Essential Roles of Cell Growth- and Apoptosis-Related Pathways in Duck Tembusu Virus Infection. Viruses, 14(10), 2225.

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RNAi silencing of LvNHE in shrimp

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Small interfering RNAs (siRNAs) were designed using the siDirect version 2.0 online program (http://sidirect2.rnai.jp/). siRNA-1, siRNA-2, and siRNA-3 designed to target LvNHE mRNA and a nontargeting siRNA (NC-siRNA) used as a negative control (Table 1) were synthesized by Sangon Biotech Company and dissolved in DEPC-H₂O. Shrimp about 1-month old with body lengths of 5.12±0.61 cm and body weights of 3.55±0.82 g were used for RNAi experiments. To confirm the interference efficiencies of the synthesized siRNAs, three siRNAs (siRNA-1, siRNA-2 and siRNA-3) were injected at the concentration of 1 μg/g body weight (bwt). In this case, DEPC-H₂O and NC-siRNA were injected as the control groups. Shrimp were cultured at 28°C in the tanks containing aerated seawater (salinity 30‰, pH 8.0). Intestines from 3 individuals in each group were respectively collected 6 and 12 h after injection. The expression levels of LvNHE were measured by real-time PCR as described above.

Li H., Ren C., Jiang X., Cheng C., Ruan Y., Zhang X., Huang W., Chen T, & Hu C. (2019). Na+/H+ exchanger (NHE) in Pacific white shrimp (Litopenaeus vannamei): Molecular cloning, transcriptional response to acidity stress, and physiological roles in pH homeostasis. PLoS ONE, 14(2), e0212887.

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Investigating USP18 Regulation in HBV

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USP18 small inhibitory RNA (siUSP18: 5′-CUGCAUAUCUUCUGGUUUATT-3′) and the negative control (NC) siRNA (NC: 5′-UUCUCCGAACGUGUCACGUTT-3′) were purchased from Sangon Biotech, China. HepAD38 cells were seeded at 3 × 10⁵/ml, 2 ml per well in 6-well plates. 24 h later, the cells were left untreated or transfected with 20 nM siUSP18 or 20 nM NC by using Lipofectamine® RNAiMAX (Invitrogen, USA) reagent according to the manufacturer’s instructions. 48 h post transfection, intracellular total protein was extracted to detect USP18 expression by western blot. Intracellular total RNA and DNA, as well as supernatant DNA, were collected respectively. Real-time PCR was performed to detect HBV pgRNA, USP18 mRNA, cccDNA and intracellular/supernatant HBV total DNA.

Li Y., Yao M., Duan X., Ye H., Li S., Chen L., Yang C, & Chen Y. (2020). The USP18 cysteine protease promotes HBV production independent of its protease activity. Virology Journal, 17, 47.

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Silencing FOXA1 in OVCAR-3 Cells

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FOXA1 siRNA and its negative control (NC) siRNA were purchased from Shanghai Sangon Co., Ltd. (Shanghai, China). Three siRNA sequences were as follows: siRNA#1 (5′-GCGACUGGAACAGCUACUATT-3′; 5′-UAGUAGCUGUUCCAGUCGCTT-3′), siRNA#2 (5′-CCACUCGCUGUCCUUCAAUTT-3′; 5′-AUUGAAGGACAGCGAGUGGTT-3′, and siRNA#3 (5′-GCACUGCAAUACUCGCCUUTT-3′; 5′-AAGGCGAGUAUUGCAGUGCTT-3′). Transfection of siRNA was performed by using Lipofectamine™ 3000 Transfection Reagent (Thermo Fisher). After transfection for 72 h, the cells were collected to evaluate the knockdown efficiency of FOXA1 in OVCAR-3 cells (n = 6) by western blot and qPCR. The siRNA with the best efficiency was used in subsequent experiments.

Wang K., Guan C., Yu J., Chen X., Shang X., Mei S., Feng X, & Zheng L. (2022). Systematic Pan-Cancer Analysis and Experimental Verification Identify FOXA1 as an Immunological and Prognostic Biomarker in Epithelial Ovarian Cancer. Disease Markers, 2022, 9328972.

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Neonatal Rat Cardiac Fibroblast Isolation and Manipulation

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Neonatal rat cardiac fibroblasts (NRCFs) were isolated from newborn SD rats (1-3 day). First, minced ventricle tissues were digested in the buffer of 40% collagenase and 60% trypsin (BioFroxx, Guangzhou, China). Then, NRCFs were separated from cardiomyocytes by gravity separation for 2 h, attached to the 10 cm cell culture dishes, and cultured with medium (DMEM+10% fetal bovine serum+1% streptomycin+1% penicillin) (Gibco, Grand Island, CA, USA). Cells were kept at 37°C in humid air containing 5% CO₂.
Cells at the second passage were exposed to miR-409-3p mimic (100 nM)/inhibitor (200 nM) versus the corresponding nc mimic/nc inhibitor (RiboBio) for 48 hours in culture medium with 1% fetal bovine serum (FBS). At the same time, cells were treated with Ang II (200 nM, 48 h, Sigma, St. Louis, MO, USA) or recombinant human TGF-β1 (20 ng/mL, 48 h, PeproTech, Rocky Hill, NJ, USA). Reagents above including Gpd1 siRNA, GATA2 siRNA, and nc siRNA (50 nM, 48 h, Sangon, Shanghai, China) were all transfected with Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's instructions.

Wang C., Yin S., Wang Q., Jiang M., Li S., Zhen W., Duan Y, & Gu H. (2022). miR-409-3p Regulated by GATA2 Promotes Cardiac Fibrosis through Targeting Gpd1. Oxidative Medicine and Cellular Longevity, 2022, 8922246.

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DTMUV Infection Modulation by IRF1 siRNA

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IFN-regulatory factor 1 siRNA (+): 5′-GCACCAGUGAUCUGUACAATT-3′, siRNA (−): 5′-UUGUACAGAUCA CUGGUGCTT -3′ and NC siRNA (+): 5′-UUCUCCGAACGUGUCACGUTT-3′, siRNA (−): 5′-ACG UGACACGUUCGGAGAATT were purchased from Sangon Biotech (Shanghai, China). Transfection of siRNA was performed using TransIntro™ EL transfection reagent according to the manufacturer’s instructions. At 6 h post-transfection, cells were infected with DTMUV at an MOI of 1 or mock-treated, and continued to incubate till harvest at 8, 16, 24, 36, and 48 hpi, respectively.

Xiang C., Huang M., Xiong T., Rong F., Li L., Liu D.X, & Chen R.A. (2020). Transcriptomic Analysis and Functional Characterization Reveal the Duck Interferon Regulatory Factor 1 as an Important Restriction Factor in the Replication of Tembusu Virus. Frontiers in Microbiology, 11, 2069.

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