Primary BMSCs were washed three times using PBS when they grew to 30%-50% fusion. Specialized media for cell transfection with low serum was used. The si-lncRNA H19 and NC with Lipofectamine® 3000 Liposome transfection reagent (Invitrogen, L3000015) were added to low-serum cell transfection-specific medium, respectively. After 8 h, a 10% FBS complete culture medium was used to replace the low serum culture medium for subsequent experiments.
Lipofectamine 3000 liposome transfection reagent
Manufactured by Thermo Fisher Scientific
Sourced in China
Lipofectamine® 3000 is a liposome-based transfection reagent designed to efficiently deliver nucleic acids, such as plasmid DNA or RNA, into a variety of mammalian cell lines. It facilitates the uptake of these molecules into the target cells, enabling gene expression studies, gene knockdown, and other molecular biology applications.
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Lab products found in correlation
4 protocols using lipofectamine 3000 liposome transfection reagent
1
Transfection of Primary BMSCs
2
Modulating Circular RNA Regulation in NSCLC
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Plasmids expressing shRNA-circ_0002360 (sh-circ_0002360), control shRNA (sh-NC), pcDNA3.1-based GPRIN1 expression plasmid (expressing the coding sequence without 3ʹUTR) GPRIN1-pcDNA, control vector, miR-585-3p mimic, mimic control miR-NC, anti-miR-585-3p and control anti-NC were gained from Kebai (Nanjing, China). For transfection, H1299/Taxol and A549/Taxol cells were plated in 12-well dishes at 1,000,000 cells per well. Next day, Lipofectamine 3000 liposome transfection reagent (Invitrogen) and the indicated plasmid (300 ng) or/and oligonucleotide (50 nM) were added into cells on the basis of the guidance of Lipofectamine 3000. 24 h later, we harvested the transfected cells for in vitro studies.
3
Lentivirus-Mediated Knockdown and Overexpression
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Cell lines were constructed using Lentivirus transduction as described previously20 (link). Lentivirus (Genechem, China) was packaged in 293T cells and transfected with Lipofectamine 3000 liposome transfection reagent (Thermo Scientific, L3000001). The interference sequence of TPPP3 as sh1:5′-CCGGGCCAATGTGGGCGTCACTAAACTCGAGTTTAGTGACGCCCACATTGGCTTTTTG-3′, sh2:5′-CCGGCTGCTCGGGTCATCAACTATGCTCGAGCATAGTTGATGACCCGAGCAGTTTTTG-3′, sh3:5′-CCGGAGGAGAGCTTCCGCAAGTTTGCTCGAGCAAACTTGCGGAAGCTCTCCTTTTTTG-3. Lentiviral constructs stabilized expressing TPPP 3 or Snail1 were obtained from Addgene. When the cells in the blank control die and the Lentivirus-transfected cells are still alive, the selected cells are expanded and sub-cultured, and cells were collected to detect the expression of the target protein.
4
Plasmid Transfection in Neuro-2a Cells
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The overexpression plasmids of ZFAS1 and MEF2C and the genetic knockout plasmids of ZFAS1 (shZFAS1, 5'-CCGTAAGTTCCCGTTTTGTGTGT-3') as well as their negative controls were synthesized by Shanghai HANBIO Company. MiR-421-3p mimic (miR10004869-1-5) and mimic control (miR1N0000001-1-5) were developed by and obtained from Guangzhou RIBIO Company. After completing the preparation of plasmid, we used the Lipofectamine 3000 liposome transfection reagent (L3000015, ThermoFisher) to transfect the overexpression/knockdown plasmid into the Neuro-2a cells of the corresponding groups. The transfection efficiency was assessed 48 h after transfection by quantitative real-time polymerase chain reaction (qRT-PCR).
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