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Transfectamine™ 7000 siRNA Transfection Reagent

HeLa cells were subjected to immunohistochemistry after transfecting with control siRNA or GAPDH siRNA using Lipofectamine 2000 and Transfectamine™ 7000. The experiment aimed to compare the efficacy of both transfection methods in knocking down GAPDH. After 48 hours of transfection, GAPDH was detected using an iFluor® 488 goat anti-mouse IgG (Cat No. 16528) through immunohistochemistry.
HeLa cells were subjected to immunohistochemistry after transfecting with control siRNA or GAPDH siRNA using Lipofectamine 2000 and Transfectamine™ 7000. The experiment aimed to compare the efficacy of both transfection methods in knocking down GAPDH. After 48 hours of transfection, GAPDH was detected using an iFluor® 488 goat anti-mouse IgG (Cat No. 16528) through immunohistochemistry.
HeLa cells were subjected to immunohistochemistry after transfecting with control siRNA or GAPDH siRNA using Lipofectamine 2000 and Transfectamine™ 7000. The experiment aimed to compare the efficacy of both transfection methods in knocking down GAPDH. After 48 hours of transfection, GAPDH was detected using an iFluor® 488 goat anti-mouse IgG (Cat No. 16528) through immunohistochemistry.
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Physical properties
SolventWater
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC12171501

OverviewpdfSDSpdfProtocol


Transfectamine™ 7000 siRNA Transfection Reagent provides a reliable and effective solution for siRNA-mediated gene knockdown experiments, efficiently delivering small interfering RNA (siRNA) and microRNA (miRNA) into diverse cell types. This reagent allows researchers to achieve high transfection efficiency with lower RNAi concentrations, resulting in effective gene knockdown with minimal non-specific effects. The low cytotoxicity of Transfectamine™ 7000 facilitates easy optimization across various concentration ranges, ensuring minimal impact on cell viability. The application protocol for Transfectamine™ 7000 is streamlined and rapid, promoting consistent and reproducible outcomes in gene silencing experiments. This reagent is capable of achieving significant levels of gene knockdown in a broad spectrum of cell lines, which is crucial for obtaining robust and conclusive experimental data. For high-throughput siRNA transfection applications, Transfectamine™ 7000 offers a straightforward workflow: the reagent is mixed with siRNA, applied to cells, incubated, and then the extent of gene knockdown is assessed. The efficiency and speed of this process, combined with the high transfection efficiency, makes Transfectamine™ 7000 an optimal choice for high-throughput screening, and it can be readily adapted for automated or robotic systems.

Example protocol


AT A GLANCE

Protocol Summary
  1. Prepare cells for transfection.

  2. Prepare Transfectamine™ 7000-siRNA mixture.

  3. Add Transfectamine™ 7000-siRNA mixture to cell culture

  4. Culture overnight.

  5. Analyze transfection efficiency with appropriate method.

Important Note

Thaw component at room temperature before starting the experiment.

PREPARATION OF WORKING SOLUTION

  1. Mix 50 to 200 nM of siRNA with 200 µL of serum-free medium.

  2. Add 7.5 µL of Transfectamine™ 7000 to Step 1.

  3. Mix well and incubate at room temperature for 20 minutes.

    Note: Ratio of Transfectamine™ 7000 and siRNA need to be optimized for different cell lines.

Sample protocol detail for 6-well and 10 cm plate
Component6 well plate (per well)10 cm plate
Fresh culture medium2 mL6 mL
siRNA
50-200 nM
50-200 nM
Serum-free medium200 µL600 µL

Transfectamine™ 7000 siRNA Transfection Reagent

~7.5 µL~22.5 µL

SAMPLE EXPERIMENTAL PROTOCOL

Preparation of Cell Culture
  1. Culture cells to ~ 90% confluency at time of transfection.

  2. Replace with fresh growth medium before transfection. For example, replace with 2 mL of medium per well for 6-well plates and 6 mL of medium for 10 cm plates.

Transtection Protocol
  1. Add Transfectamine™ 7000-siRNA mixture to the culture plate and culture overnight.

    Note: Recombinant protein can start to be detected as early as 16 hours post-transfection. Maximal expression level may be observed 72~96 hours post-transfection.

Images


References


View all 50 references: Citation Explorer
Rational polyelectrolyte nanoparticles endow preosteoclast-targeted siRNA transfection for anabolic therapy of osteoporosis.
Authors: Zhang, Zheng and Ding, Peng and Meng, Yichen and Lin, Tao and Zhang, Zhanrong and Shu, Haoming and Ma, Jun and Cohen Stuart, Martien and Gao, Yang and Wang, Junyou and Zhou, Xuhui
Journal: Science advances (2023): eade7379
Self-Assembled Oleic Acid-Modified Polyallylamines for Improved siRNA Transfection Efficiency and Lower Cytotoxicity.
Authors: Salvador, Cristian and Andreozzi, Patrizia and Romero, Gabriela and Loinaz, Iraida and Dupin, Damien and Moya, Sergio E
Journal: ACS applied bio materials (2023): 529-542
Bioresponsive Chimaeric Polymersomes Mediate Sustained and Liver-Specific siRNA Transfection In Vivo.
Authors: Huang, Ri and Wang, Feifei and Fu, He and Qi, Xinming and Xing, Guozhen and Ren, Jin and Cheng, Liang and Meng, Fenghua and Zhong, Zhiyuan
Journal: Biomacromolecules (2023): 5353-5363
A Bayesian Implementation of Quality-by-Design for the Development of Cationic Nano-Lipid for siRNA Transfection.
Authors: Bastogne, Thierry and Hassler, Lucie and Bruniaux, Jonathan and Thomassin, Magalie and Gidrol, Xavier and Sulpice, Eric and Navarro, Fabrice P
Journal: IEEE transactions on nanobioscience (2023): 455-466
Two-Photon Light Trigger siRNA Transfection of Cancer Cells Using Non-Toxic Porous Silicon Nanoparticles.
Authors: Chaix, Arnaud and Cueto-Diaz, Eduardo and Dominguez-Gil, Sofia and Spiteri, Chantelle and Lichon, Laure and Maynadier, Marie and Dumail, Xavier and Aggad, Dina and Delalande, Anthony and Bessière, Aurélie and Pichon, Chantal and Chiappini, Ciro and Sailor, Michael J and Bettache, Nadir and Gary-Bobo, Magali and Durand, Jean-Olivier and Nguyen, Christophe and Cunin, Frédérique
Journal: Advanced healthcare materials (2023): e2301052
Cationic amino-acid functionalized polymethacrylamide vectors for siRNA transfection based on modification of poly(2-isopropenyl-2-oxazoline).
Authors: Jerca, Florica Adriana and Muntean, Cristina and Remaut, Katrien and Jerca, Valentin Victor and Raemdonck, Koen and Hoogenboom, Richard
Journal: Journal of controlled release : official journal of the Controlled Release Society (2023): 687-699
siRNA Transfection Mediated by Chitosan Microparticles for the Treatment of HIV-1 Infection of Human Cell Lines.
Authors: Chronopoulou, Laura and Falasca, Francesca and Di Fonzo, Federica and Turriziani, Ombretta and Palocci, Cleofe
Journal: Materials (Basel, Switzerland) (2022)
Small Interfering RNA (siRNA) Transfection in Epiblast Stem Cells.
Authors: Kafer, Georgia R
Journal: Methods in molecular biology (Clifton, N.J.) (2022): 47-55
The Characteristics of Human iPS Cells and siRNA Transfection Under Hypoxia.
Authors: Sugimoto, Kouji and Hayashi, Yoshihiko
Journal: Methods in molecular biology (Clifton, N.J.) (2022): 657-664
Non-viral siRNA transfection of primary mesenchymal stromal cells (MSCs): Assessment of tyrosine-modified PEI and PPI efficacy and biocompatibility.
Authors: Noske, Sandra and Karimov, Michael and Hansen, Max and Zatula, Nathalie and Ewe, Alexander and Aigner, Achim
Journal: International journal of pharmaceutics (2022): 121359