DNAJC3-AS1 and miR-140 were overexpressed in HL60, and U937 cells by transfecting DNAJC3-AS1 vector (pcDNA3.1 backbone) and/or miR-140 mimic (RiboBio, China). All transfections were mediated by Neon Electroporation Transfection device (Thermo Fisher Scientific). Negative control (NC) experiments were performed, transfecting cells with empty vector or NC miRNA. Untransfected cells were cultivated until the following assays to serve as the control. The overexpression was checked every 24 h until 96 h.
Neon electroporation transfection device
Manufactured by Thermo Fisher Scientific
Sourced in China
The Neon Electroporation Transfection device is a laboratory equipment used for the delivery of nucleic acids, such as DNA or RNA, into cells. It utilizes electroporation technology to temporarily create pores in the cell membrane, allowing the uptake of the desired genetic material. The device provides controlled electrical parameters to optimize the transfection efficiency for various cell types.
Automatically generated - may contain errors
Lab products found in correlation
Mir 140 mimic, by RiboBio (1 mentions)
Nucleospin rna plus xs, by Takara Bio (1 mentions)
Mda mb 436, by American Type Culture Collection (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Penicillin, by Merck Group (1 mentions)
Streptomycin, by Merck Group (1 mentions)
4 protocols using neon electroporation transfection device
1
Overexpression of DNAJC3-AS1 and miR-140 in Leukemia Cells
2
DNAJC3-AS1 and miR-27b Transfection in HepG2 and Huh7 Cells
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HepG2 and Huh7 cells were transfected with DNAJC3-AS1, si-DNAJC3-AS1, or miR-27b mimic using Neon Electroporation Transfection device (Thermo Fisher Scientific) according to the manufacturer’s protocol. Cells cultured in fresh medium without transfections until the end of experiment were served as the control (C). The sequence of MiR-27b mimics was (5ʹ-TTCACAGTGGCTAAGTTCTGCAA-3ʹ), and the target sequences of DNAJC3-AS1 siRNA were ggugcugaauguggaguaatt (F) and uuacuccacauucagcacctt (R).
NucleoSpin RNA Plus XS (Takara Bio) was used to isolate total RNA from paired tissues and cells of HepG2 and Huh7 cell lines. To reduce DNA contamination, all RNA samples were incubated with DNase I (NEB) to achieve a ratio of OD260/280 above 1.8. Bioanalyzer was used to make sure all RNA samples were with a RIN value higher than 8.0.
NucleoSpin RNA Plus XS (Takara Bio) was used to isolate total RNA from paired tissues and cells of HepG2 and Huh7 cell lines. To reduce DNA contamination, all RNA samples were incubated with DNase I (NEB) to achieve a ratio of OD260/280 above 1.8. Bioanalyzer was used to make sure all RNA samples were with a RIN value higher than 8.0.
3
TNBC Cell Line Culture and DRAIR Overexpression
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Human TNBC cell lines MDA-MB-436 and SUM149PT from ATCC were cultured in DMEM medium (Cat # 11,054,001, Thermo Fisher Scientific, Shanghai, China) supplemented with 10% FBS (Cat # MFCD00132239, Sigma-Aldrich, Shanghai, China), streptomycin (10 μg/ml, Cat # 3810–74-0, Sigma-Aldrich, Shanghai, China) and penicillin (100 U/ml, Cat # 87–08-1, Sigma-Aldrich, Shanghai, China) in an incubator at 37°C with 95% humidity and 5% CO2.
MDA-MB-436 (ATCC, USA) and SUM149PT (ATCC, USA) cells were overexpressed with DRAIR by transfecting cells with DRAIR expression vector (pcDNA3.1) using Neon Electroporation Transfection device (Thermo Fisher Scientific, Shanghai, China). Transected cells were cultured in fresh media prior to subsequent assays.
MDA-MB-436 (ATCC, USA) and SUM149PT (ATCC, USA) cells were overexpressed with DRAIR by transfecting cells with DRAIR expression vector (pcDNA3.1) using Neon Electroporation Transfection device (Thermo Fisher Scientific, Shanghai, China). Transected cells were cultured in fresh media prior to subsequent assays.
4
Overexpression of circ-MYBL2 and miR-19a in SW1990 and PANC-1 cells
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SW1990 and PANC-1 cells were overexpressed with circ-MYBL2 and/or miR-19a through the transfection of circ-MYBL2 (pcDNA3.1) expression vector and/or mimic of miR-19a (RiboBio). Transfection was performed with Neon Electroporation Transfection device (Thermo Fisher Scientific). In each transfection, 106 cells were transfected with either 10 µg vector or 50 nM miRNA. After transfection, cells were subjected to cell culture in fresh medium for a total of 48 h prior to the subsequent assays.
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