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Rpa Ppt Template Lipofectamine®3000 Transfection Reagent Protocol Transfect cells according to the following table. Use the indicated volume of DNA and P3000™Reagent with each of the two volumes of Lipofectamine®3000 (when performing optimization). Each reaction mix volume is for one well and accounts for pipetting variations.

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Transfection Protocol Calculator › Our Lipofectamine Reagent Protocols have been optimized for efficiency, viability, and reproducibility across a broad range of cell types. This is often the best place to start, especially in a new cell line.
Each cell line was transfected with a luciferase reporter plasmid using either ViaFect™ Transfection Reagent, Lipofectamine® 2000 Transfection Reagent or Lipofectamine® 3000 Transfection Reagent following manufacturer instructions, applying either 5µl (50ng of DNA) or 10µl (100ng of DNA) of the transfection complex in each well of a 96 ... LipofectAMINE 148.1 µL 200 nM, Vt = 3200 µL add 2900 L Opti-MEM 25 nM, Vt = 2000 µL 500 µL 100 nM solution 1500 µL Opti-MEM dispense 200 µL/well overnight transfection complex formation 100 nM, Vt = 3000 µL 1500 µL 200 nM solution 1500 µL Opti-MEM Opti-MEM Opti-MEM Figure 4.13.2 Preparation of LipofectAMINE-LNA complexes for cellular ...

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Lipofectamine 3000 is the way to go! 😁 jk I haven’t tried Calcium phosphate so can’t comment on that. Lipofectamine 3000 reagent provided higher GFP transfection efficiency than Lipofectamine 2000 and FuGENE HD reagents for all five cell lines. Figure 2. The transfection protocol for Lipofectamine 3000 was developed to be easy to use while still ensuring optimum performance and reliability in a wide panel of cell lines. Lipofectamine 2000 is efficient in transfecting DNA into HEK293 cells. ... I have been using Lipofectamine™ 3000 Transfection Reagent to transfect human and mouse ...

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We tested 8 commercial transfection reagents (Lipofectamine 3000, Lipfectamine Stem, EndoFectin, TransFectin, JetPRIME, Transfex, Effectene, PEI-NLS Peptide) with 4 different concentrations each and eGFP plasmid. Ex: conditions for Lipofectamine 3000 were All reagents were used according to manufacturer’s protocol. Original Method Objective 2
In IB3–1 monolayers, LipofectAMINE PLUS and LipofectAMINE 2000 alone or together with Effectene transfected monolayers effectively so that CFTR protein could be detected by immunoprecipitation (Fig. 8). Effectene and FuGENE 6.0 alone or in combination failed to transduce IB3–1 monolayers efficiently enough to observe a CFTR protein signal. Transfection is a valuable tool for inserting genes of interest into cells. To measure transfection efficiency, often a reporter gene such as GFP is co-expressed with the gene of interest. The NucleoCounter ® NC-3000™ comes equipped with a GFP-transfection efficiency assay that is quick and easy and uses very little sample volume.

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Lipofectamine 3000 reagent 2. Electroporation Posttransfection, bright-field Posttransfection, GFP: 57% transfection efficiency . ... Optimization of transfection effi ciency, depending on the experimental need of the researcher, can be achieved by modulating plating density, with minimal effect on cell ...
Virtual Screening and Optimization of Novel mTOR Inhibitors for Radiosensitization of Hepatocellular Carcinoma. Feng YQ, Gu SX, Chen YS, Gao XD, Ren YX, Chen JC, Lu YY, Zhang H, Cao S. Drug Design, Development and Therapy 2020, 14:1779-1798 Published Date: 8 May 2020 Lipofectamine™ 3000 Transfection Reagent leverages our most advanced lipid nanoparticle technology to provide superior transfection performance with improved application outcomes and reproducible results.

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In Experiment 1, using 96 well-plate and a robotic inverted fluorescent microscope, we compared transfection efficiency among commercially available transfection reagents (TR) Lipofectamine® 3000 (Lipo3), Lipofectamine® LTX (LipoLTX), and TransIT-X2® (TransX2), three doses of TR (i.e., 0.15, 0.3, and 0.4μL/well), and three doses of Green ...
Transfections were conducted in a 24-well plate with 300 ng pX330, 400 ng pEF-BFP-Puro R, 50 ng sgRNA vector, 1.5 μl Lipofectamine 3000 (ThermoFisher Scientific) and 1 μl P3000 transfection reagent. Cells were passaged at 72 h post-transfection into a single well of a 6-well plate and selected with 0.5 μg/ml puromycin for 2 weeks. Lipofectamine 3000 reagent maintains a high transfection efficiency within a robust dynamic range of lipid doses for quick and easy optimization. The effect of the compound was even more pronounced in transfections with linear forms of polyoma DNA, suggesting that chloroquine inhibits degradation of DNA absorbed by the cells.

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Optimization Protocols. ... transfection efficiencies with TransIT®-2020 and TransIT®-LT1 were clearly higher than with Lipofectamine® 3000. Fedir Kiskin.
Lipofectamine 3000 (Invitrogen) was used for cell transfection following the protocol. Stable ovarian cancer cells overexpressing -193a-3p were miR harvested after 14 days of G418 selection and verified by QPCR. Ovarian cancer cells with stable GRB7 knockdown were infected with lentiviral particles carrying GRB7 shRNA plasmid (Santa Cruz). After Lipofectamine 3000 reagent transfection efficiency Protein expression improvement (x-fold), Lipofectamine 3000 vs. Lipofectamine 2000 reagents 3T3 4 4T1 2 A431 2 A549 3 ACHN 2 bEnd.3 9 BJ 3 BT-549 4 C2C12 3 C6 5 Caco-2 2 Caki-1 4 CHO-K1 1 CHO-S 1 COLO 205 4 COS-7 4 DU 145 2 H460 3 H9c2 3 HCC1937 5 HCT116 1 Cell type Lipofectamine 3000 reagent ...

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transfection reagent Lipofectamine® 3000. We describe this new technology and how this simple versatile reagent delivers efficient, reporducible transfection of both DNA and RNA ino a wide range of hard-to-transfect, biologically relevant cell models. In the present study, we demonstrated that Lipofectamine® 3000 can be used to Lipofectamine® 3000 Reagent per well 0.15 and 0.3 μL 0.75 and 1.5 μL 3.75 and 7.5 μL Transfection of siRNA To transfect cells with siRNA, follow the protocol as described for DNA but do not add P3000™ Reagent when diluting the siRNA (step 3). Limited Product Warranty

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We used 100-300 ng of DNA per unique plasmid with 1 ul of P3000 reagent and 1 ul of Lipofectamine 3000, all prepared in 100 ul Opti-MEM per 24-well plate. In the 24-well plate format, 200 000 HEK293FT cells or 150 000 HeLa cells were transfected per well.
The methodology follows the manufacturer’s protocol with some modifications. The transfection procedure was carried out using SOX-trio/DNA (2.5 ng/µL and 5.0 ng/µL) and Lipofectamine reagents with different ratios or volumes. The cells were cultured and transfected using DNA-to-lipofectamine ratio of 1:1, 2:1 and 2:2.