Small interference RNA targeting DLX6-AS1 (si-DLX6-AS1: 5ʹ-AAUAAAGAACACUUACACUACUG-3ʹ) together with its negative control (si-NC: 5ʹ- UUCUCCGAACGUGUCACGUTT-3ʹ) were assembled by Ribobio (Guangzhou, China). miR-506-3p mimics (miR-506-3p; Product ID: miR10002878-1-5), miR-506-3p inhibitors (anti-miR-506-3p; Product ID: miR20002878-1-5) and respective controls, including miR-NC (Product ID: miR1N0000001-1-5) and anti-miR-NC (Product ID: miR2N0000001-1-5) were purchased from Ribobio. The sequences of DLX6-AS1 and STAT2 were amplified and cloned into the pcDNA3.1 overexpression vector, named as pcDNA-DLX6-AS1 and pcDNA-STAT2 (Sangon, Shanghai, China), and pcDNA empty vector was served as the negative control (pcDNA-NC). For stable DLX6-AS1 knockdown, a lentiviral vector containing short hairpin RNA targeting DLX6-AS1 (sh-DLX6-AS1: 5ʹ-GGTTCAGTATAGATTTCTA-3ʹ) and its negative control (sh-NC: 5ʹ- TTCTCCGAACGTGTCACGT-3ʹ) were synthesized by Research-Bio Co., Ltd (Shanghai, China). These oligonucleotides (20 nM) and plasmids (1 µg) were transfected into SK-N-SH and LAN-6 cells (5×104 cells) using Lipofectamine 3000 Reagent (Invitrogen, Carlsbad, CA, USA).
Pcdna nc
Manufactured by Sangon
Sourced in China
PcDNA-NC is a laboratory plasmid vector. It is commonly used for the expression and production of recombinant proteins in mammalian cell lines.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (4 mentions)
Lipofectamine 3000 reagent, by Thermo Fisher Scientific (3 mentions)
Pcdna dlx6 as1, by Sangon (2 mentions)
Anti mir nc, by Sangon (2 mentions)
Si rp11 59j16, by Sangon (2 mentions)
Si mcm2, by Sangon (2 mentions)
Pcdna mcm2, by Sangon (2 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (2 mentions)
Si nc, by Sangon (2 mentions)
Mimic nc, by Sangon (2 mentions)
Anti mir nc, by RiboBio (1 mentions)
Mir 506 3p, by RiboBio (1 mentions)
Anti mir 506 3p, by RiboBio (1 mentions)
Si nc, by RiboBio (1 mentions)
Mir nc, by RiboBio (1 mentions)
Sh nc, by Sangon (1 mentions)
Lipofectamine 3000 transfection reagent, by Takara Bio (1 mentions)
Pcdna, by Sangon (1 mentions)
Mir nc, by Sangon (1 mentions)
Anti mir 760, by Sangon (1 mentions)
13 protocols using pcdna nc
1
Targeting DLX6-AS1 and miR-506-3p in Neuroblastoma
2
Transfection Optimization for LINC01001, IGF2BP2, and MYC
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The transfection dose for sh-LINC01001, sh-IGF2BP2, sh-MYC, pcDNA- LINC01001 plasmids and its negative control sh-NC and pcDNA-NC (synthesized by Sangon Biotech, Shanghai, China) were 2 μg for A549/R and H1299/R cells in each well of 6-well plates. All the transfection was performed using Lipofectamine™ 3,000 Transfection Reagent (Takara, Kusatsu, Japan). Following 48 h transfection, A549/R and H1299/R cells were applied to subsequent experiments.
3
Investigating circYIPF6-PTBP1 Axis in Glioma
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The shRNA targeted circYIPF6 (sh-circYIPF6) and non-sense sh-NC were constructed by RIBOBIO (Guangzhou, China). miR-760 mimics (miR-760), miR-760 inhibitors (anti-miR-760), and their control mock (miR-NC, anti-miR-NC), together with pcDNA3.1-PTBP1 and pcDNA3.1 control vector (pcDNA-NC) were offered by Sangon Biotech (Shanghai, China) and then transfected glioma cells for 48 h.
4
Overexpressing NOX2 and Modulating miR-126a-5p
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The empty vector (pcDNA-NC) and NOX2-overexpressing vector (pcDNA-NOX2) were from Sangon Biotech (Shanghai, China). The miR-126a-5p mimic and inhibitor for functional gain or reduction, respectively, were obtained from RiboBio (Guangzhou, China). For cell transfection, miR-126a-5p mimic, miR-126a-5p inhibitor, or pcDNA-NOX2 were mixed with transfection agent Lipofectamine 2000. After 20 min, the mixture was added into the cultured cells in 24-well plates to reach a concentration of 100 nmol/well. Following the transfection for 6 h, cells were cultured for another 48 h in normal media and collected for measurement of enzyme activity, protein, mRNA, and others.
5
Modeling Alzheimer's in SH-SY5Y Cells
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Human neuroblastoma cell line (SH-SY5Y) was purchased from Beina Cell Bank and cultured in DMEM/F12 complete medium containing 10% fetal bovine serum and 1% antibiotics. The cells were cultured in an incubator containing 5% CO2, 95% humidity and 37°C. SH-SY5Y was treated with different concentrations (0, 5, 10, or 20 µM) of Aβ for 24 h or 10 µM of Aβ for different times (0, 12, 24, or 48 h) to establish in vitro AD cell model. si-RP11-59J16.2, si-RNA negative control (si-NC), si-MCM2, pcDNA-MCM2, pcDNA-NC were all purchased from Sangon Biotech (Shanghai) Co., Ltd. The antibody was purchased from Abcam. CCK-8 cell viability detection kit and Annexin V-FITC/PI Cell Apoptosis Assay kit were purchased from BEENbio Biotechnology.
6
HOXA-AS2 Regulation in Tca-8113 Cells
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The short-hairpin RNA (shRNA) against HOXA-AS2 (shRNA-HOXA-AS2-1 and shRNA-HOXA-AS2-2) and corresponding empty vector (shRNA-NC) were purchased from GenePharma. The pcDNA-EZH2 and the empty vector pcDNA-NC were constructed from Sangon Biotech. Tca-8113 cells (1 × 106 cells/well) were inoculated into 12-well plates and then transfected with shRNA-NC, shRNA-HOXA-AS2, pcDNA-NC, and pcDNA-EZH2 using Lipofectamine® 3000 reagent (Thermo Fisher Scientific) following the manufacturer’s instruction. After 48 h, RT-qPCR was used to assess the transfection efficiencies. The transfected cells were collected for the next experiments.
7
Modulating DLBCL via miR-125b-5p and TNFAIP3
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According to the manufacturer's instructions, miR-125b-5p inhibitor and its negative control (NC) (50 nM, GenePharma Co., Ltd.), pcDNA-TNFAIP3, and pcDNA-NC (40 nM, Sangon Biotech Co., Ltd.) vectors were transfected into LY8 or DUL cells using Lipofectamine™ 2000 (Thermo Fisher Scientific) according to the manufacturer's instructions. The DLBCL cells were then incubated with the conditioned medium from SUD cells treated with 30 µg EVs or GW4869 for 24 h at 37°C.
miR-125b-5p inhibitor and its NC were transfected into SUD cells using Lipofectamine™ 2000, followed by incubation for 4 h at 37°C. The cells were cultured in complete medium for 48 h, and then EVs were isolated and incubated with LY8 and DUL cells.
miR-125b-5p inhibitor and its NC were transfected into SUD cells using Lipofectamine™ 2000, followed by incubation for 4 h at 37°C. The cells were cultured in complete medium for 48 h, and then EVs were isolated and incubated with LY8 and DUL cells.
8
Transfection and Post-Transfection Assays
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pcDNA‐ARPP21, pcDNA‐NC, mimic NC, and miR‐128 mimic were purchased from Sangon Biotech (Shanghai, China). The cell transfection was performed using Lipofectamine™ 2000 (Invitrogen, Carlsbad, CA, USA). The subsequent experiments were conducted after 48 h.
9
Targeting circTM7SF3 with siRNAs in immune cells
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CircTM7SF3 specific small interfering RNAs (100 nM), including si-circTM7SF3_1 (5′-UGGAUUUGUACCAUUCUUCUG-3′), si-circTM7SF3_2 (5′-ACUCAUUGGUUCCUUUAAGGG-3′) and si-circTM7SF3_3 (5′-UCAUAUUCUGAAUCUCAUCCU-3′), siRNA negative control (si-NC; 100 nM), miR-206 (40 nM), miR-NC (40 nM), miR-206 inhibitor (anti-miR-206; 20 nM), anti-miR-NC (20 nM), ASPH ectopic expression plasmid (pcDNA-ASPH; 1 μg) and pcDNA-NC (1 μg) from Sangon Biotech were transfected into differentiated THP-1 cells or differentiated hMDMs in 6-well plates using Lipofectamine® 2000 reagent (Invitrogen; Cat. no. 11668019).
10
miRNA-mediated regulation of vascular smooth muscle cells
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hVSMC was incubated in a 24-well plate in advance until the cell confluence was up to about 40%. Commercial RNA transfection reagent EntransterTM-R4000 (Engreen Biosystem, China) was applied to transfect miRNA mimics or sh-RNA. MiR-29b-3p mimics, NC mimics, pcDNA-IGF1 and pcDNA-NC were synthesised by Sangon Biotech. Specifically, the miRNA mimics or pcDNA were diluted in a serum-free medium, then mixed with the diluted transfection reagent, and added to the well. After 48 h, subsequent experiments could be carried out.
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