The miR-19a-3p inhibitor (inhibitor) and its negative control (miR-NC) were synthesized by GenePharma Co., Ltd. (Shanghai, China). The pcDNA3.1/IGFBP3 vector was generated by inserting the open reading frame of IGFBP3 without 3′UTR into the pcDNA3.1 vector (Sangon Biotech, Shanghai, China). Small interference RNA against IFGBP3 (si-IGFBP3) was purchased from GenePharma Co., Ltd. Before OGD/R, SH-SY5Y cells were seeded at a density of 3 × 105 cells per well and transfected with inhibitor, miR-NC, IGFBP3 or empty vector. In the rescue experiments, si-IGFBP3 was transfected into miR-19a-3p silenced SH-SY5Y cells. All these transfection procedures were performed for 48 h using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) following the manufacturer’s protocols.
Mir nc
Manufactured by GenePharma
Sourced in China, United States, Puerto Rico
The MiR-NC is a laboratory equipment product developed by GenePharma. It is designed to perform the core function of small RNA normalization control. The product provides a standardized solution for normalizing small RNA expression data obtained from experimental analysis.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (197 mentions)
Si nc, by GenePharma (118 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (107 mentions)
Anti mir nc, by GenePharma (96 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (70 mentions)
Lipofectamine 3000 reagent, by Thermo Fisher Scientific (50 mentions)
Sh nc, by GenePharma (42 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (40 mentions)
Mir nc, by Thermo Fisher Scientific (37 mentions)
Anti nc, by GenePharma (25 mentions)
Pcdna3, by Thermo Fisher Scientific (21 mentions)
Inhibitor nc, by GenePharma (20 mentions)
Pcdna3.1 vector, by Thermo Fisher Scientific (19 mentions)
In mir nc, by GenePharma (18 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (16 mentions)
Pcdna nc, by GenePharma (14 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (14 mentions)
Lipofectamine 2000 transfection reagent, by Thermo Fisher Scientific (14 mentions)
Pcdna3.1 vector, by GenePharma (8 mentions)
Si nc, by Thermo Fisher Scientific (8 mentions)
532 protocols using mir nc
1
Modulating miR-19a-3p and IGFBP3 in SH-SY5Y cells
2
Modulating miRNA-150-5p and VDR in Fibroblasts
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miRNA-150-5p mimics and negative controls (miR-NC) were designed and synthesized by Shanghai GenePharma Co., Ltd. as follows: miR-150-5p mimics forward, 5'-UCUCCCAACCCUUGUACCAGUG-3' and reverse, 5'-CACUGGUACAAGGGUUGGGAGA-3' and miR-NC forward, 5'-UCACAACCUCCUAGAAAGAGUAGA-3' and reverse, 5'-UCUACUCUUUCUAGGAGGUUGUGA-3'. Plasmid for vitamin D receptor (VDR) overexpression was constructed by inserting the amplified complementary DNA (cDNA) into pcDNA 3.1 vector (Invitrogen; Thermo Fisher Scientific, Inc.). Fibroblasts were incubated at 37˚C overnight in antibiotics-free DMEM medium until 70% confluence, and transfection was performed with 50 nM oligonucleotides or 2 µg vectors using Lipofectamine® 2000 (Invitrogen; Thermo Fisher Scientific, Inc.) according to the manufacturer's instructions for 4-6 h at 37˚C before changing to DMEM medium. Following 48 h transfection, fibroblasts were stimulated at 37˚C with 50 BMP-2 and 10 ng/ml TGF-β1 for 14 days before further analysis.
3
Retinoblastoma Cell Lines Transfection
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Both the RB cell lines (SO-RB50, WERI-Rb1, Y79 and RBL-13) and the healthy human retinal epithelial cell line (ARPE-19) were acquired from the American Type Culture Collection (Shanghai, China). They were maintained in RPMI-1640 media (Gibco, Carlsbad, CA) with 10% fetal bovine serum (FBS) (Invitrogen, Carlsbad, CA) at 37°C in a humidified environment of 95% air and 5% CO2. siRNAs targeting TP53TG1 (si-TP53TG1), negative control siRNA (si-NC), miR-33b mimics, NC mimics (miR-NC), miR-33b inhibitors, and corresponding NC (miR-NC) were acquired from GenePharma (Shanghai, China). Transient transfections were performed in RB cells using lipofectamine 3000 (Invitrogen, Carlsbad, CA), following manufacturers’ protocols.
4
Silencing KIFC1 and Overexpressing miR-338-3p
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The small interference RNA (siRNA) sequences were designed for KIFC1: sense 5′-UCG AAA UGA GAA AUC UCG GAG-3′, antisense 5′-CCG AGA UUU CUC AUU UCG AAU-3′. The negative control siRNA (siRNA-NC) were random sequences that have no homology with any known mammalian gene. KIFC1 siRNA (si-KIFC1) and negative control (siRNA-NC) were synthesized by GenePharma Company (Shanghai, P.R. China). To overexpress miR-338-3p, miR-338-3p mimic and mimic negative control (miR-NC) were purchased from GenePharma. 786-O, 769-P, and OS-RC-2 cells were seeded at 1 × 105 cells per well in six-well plates and incubated overnight at 37°C with 5% CO2. Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) was used to transiently transfect cells with si-KIFC1, siRNA-NC, miR mimic, or miR-NC (GenePharma). The efficiency of transfection was evaluated by real-time quantitative PCRRT-qPCR and Western blot analysis after 48 h of transfection.
5
miR-1 Overexpression and Knockdown in Gastric Cancer
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miR-1 mimics, scramble miRNA negative control (miR-NC) and miR-1 inhibitors were synthesized by Shanghai GenePharma Co., Ltd.. The sequences were as follows: miR-1 mimic, 5′-UGGAAUGUAAAGAAGUAUG UAU-3′ (sense) and 5′-ACAUACUUCUUUACAUUCCAUU-3′ (antisense); miR-NC, 5′-UUCUCCGAACGUGUCACGUTT-3′ (sense) and 5′-ACGUGACACGUUCGGAGAATT-3′ (antisense); miR-1 inhibitor, 5′-UUCAGUUAUCACAGUACU GUA-3′; inhibitor NC, 5′-CAGUACUUUUGUGUAGUA CAA-3′. SGC7901, SGC7901/ADM and SGC7901/VCR cells were transfected with 100 nM miR-1 mimics or miR-1 inhibitor for miR-1 overexpression and knockdown, respectively. miR-NC (100 nM) and inhibitor NC (100 nM) were used as negative controls. Transfection was conducted using Lipofectamine 2000 reagent (Invitrogen; Thermo Fisher Scientific, Inc.) according to the manufacturer's protocol. Six hours post-transfection, the medium was changed with fresh medium. At 48 h post-transfection, the cells were used in subsequent experiments.
6
Modulating miR-130b and CYLD Expression
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The agonist and antagonist of miR-130b (miR-130b(+) and miR-130b(–)) and their negative controls (miR-NC(+) and miR-NC(–)) were designed and synthesized by GenePharma Company (China). The silence vector of CYLD (sh-CYLD) and its negative control (sh-NC) were also designed and synthesized by GenePharma Company. All vectors and microRNAs were transfected using Lipofectamine 2000 (Thermo Fisher Scientific, USA) in accordance with instructions.
7
miR-195-5p Regulation of E2F3 in AMC-HN-8 Cells
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miR-195-5p mimic and the negative control miRNA (miRNA-NC) were obtained from Guangzhou RiboBio Co., Ltd. The E2F3 overexpression vector pcDNA-E2F3 and empty control vector pcDNA-NC were constructed by Shanghai GenePharma Co., Ltd. The sequences were as follows: miR-195-5p mimic sense, 5'-UAGCAGCACAGAAAUAUUGGC-3'; miR-195-5p mimic antisense, 5'-CAAUAUUUCUGUGCUGCUAUU-3'; mimics negative control (miR-NC) sense, 5'-UUCUCCGAACGUGUCACGUTT-3'; miR-NC antisense, 5'-ACGUGACACGUUCGGAGAATT-3'. Cells were plated into 6-well plates (1x106 cells per well), and transfection was performed when cells at the logarithmic growth phase reached 80% confluence. According to the product instructions, AMC-HN-8 cells were transfected with miR-195-5p mimic (50 nM), miR-NC (50 nM), pcDNA-E2F3 (100 nM) and pcDNA-NC (100 nM) using Lipofectamine® 2000 (Invitrogen; Thermo Fisher Scientific, Inc.). The transfection efficiencies were assessed by RT-qPCR at 48 h after transfection.
8
Modulation of miR-148a-3p in Macrophages
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RAW264.7 cells were randomly divided into NC (miR mimic) group, miR mimics group, NC (miR inhibitor) group, miR inhibitor group, LPS + NC (miR mimic) group, LPS + NC (miR inhibitor) group, LPS+ miR inhibitor group and LPS + miR mimics. The MiR-148a-3p mimic, miR-148a-3p inhibitor and their negative controls were transfected into RAW264.7 macrophages using Lipofectamine 2000 Transfection Reagent (Invitrogen, USA) according to the manufacturer's instructions. Twenty-four hours after transfection, cells were used for further experiments. MiR-148a-3p mimic and its negative control miR-NC, miR-148a-3p inhibitor and its negative control miR-NC were purchased from Shanghai GenePharma Co. The cells were collected 4 h later for LPS (1 μg/mL) stimulation for RT-PCR and 24 h for western blotting.
9
Silencing AGAP2-AS1 and TGF-β1 in GBM
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In short, the oligonucleotides (RiboBio, Guangzhou, China): si-AGAP2-AS1: 5′-AUAAAGCAGGUAACAAGUGGG-3’ (sense), 5′-CACUUGUUACCUGCUUUAUAA-3′(antisense); si-TGF-β1: 5′-ACGGAAAUAACCUAGAUGGGC-3’ (sense), 5′-CCAUCUAGGUUAUUUCCGUGG-3′(antisense), miR-486-3p mimic/inhibitor (miR-486-3p/miR-486-3p in) and their controls (si-NC, miR-NC, and miR-NC in), and plasmids (GenePharma, Shanghai, China): pcDNA and pcDNA-TGF-β1 (TGF-β1, NM_000660.7), were collected in this research. Using Lipofectamine 3000 (Invitrogen, #L3000075 ), 50 nM oligonucleotides and 6 μg plasmids were transfected into GBM cells at 70 % confluence for 48 h, followed by the assessment of the transfection efficiency.
10
Modulating miR-1270 and CDR1as Expressions
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The RNA oligoribonucleotides used in this study, including miR-1270 mimic, miR-1270 inhibitor, the small interfering RNAs (siRNAs) targeting CDR1as (si-CDR1as) or SCAI (si-SCAI), and the corresponding miRNA control (miR-NC) and siRNA control (si-NC), were purchased from GenePharma (Shanghai, China). The RNA oligoribonucleotides used in this study, including miR-1270 mimic, miR-1270 inhibitor, the siRNAs targeting CDR1as (si-CDR1as) or SCAI (si-SCAI), and the corresponding miRNA control (miR-NC) and siRNA control (si-NC), were purchased from GenePharma (Shanghai, China).
The lentivirus targeting human CDR1as was purchased from GeneChem (Shanghai, China). The targeting sequence was designed as follows: 1 sense 5′-TGCACCTGTGTCAAGGTCTTTTCAAGAGAAAGACCTTGACACAGGTGCTTTTTTC-3′ and 2 sense 5′-TGGTCTTCCAGCGACTTCAATTCAAGAGATTGAAGTCGCTGGAAGACCA-3′. miR-7 mimics and inhibitors were synthesized by RiboBio. The miRNA oligonucleotides were transfected using Lipofectamine RNAiMAX (50 nmol/L; Invitrogen, CA, USA).
The lentivirus targeting human CDR1as was purchased from GeneChem (Shanghai, China). The targeting sequence was designed as follows: 1 sense 5′-TGCACCTGTGTCAAGGTCTTTTCAAGAGAAAGACCTTGACACAGGTGCTTTTTTC-3′ and 2 sense 5′-TGGTCTTCCAGCGACTTCAATTCAAGAGATTGAAGTCGCTGGAAGACCA-3′. miR-7 mimics and inhibitors were synthesized by RiboBio. The miRNA oligonucleotides were transfected using Lipofectamine RNAiMAX (50 nmol/L; Invitrogen, CA, USA).
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