Transfectamine™ 7000 siRNA Transfection Reagent
Example protocol
AT A GLANCE
Prepare cells for transfection.
Prepare Transfectamine™ 7000-siRNA mixture.
Add Transfectamine™ 7000-siRNA mixture to cell culture
Culture overnight.
Analyze transfection efficiency with appropriate method.
Thaw component at room temperature before starting the experiment.
PREPARATION OF WORKING SOLUTION
Mix 50 to 200 nM of siRNA with 200 µL of serum-free medium.
Add 7.5 µL of Transfectamine™ 7000 to Step 1.
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.
Component | 6 well plate (per well) | 10 cm plate |
Fresh culture medium | 2 mL | 6 mL |
siRNA | 50-200 nM | 50-200 nM |
Serum-free medium | 200 µL | 600 µL |
Transfectamine™ 7000 siRNA Transfection Reagent | ~7.5 µL | ~22.5 µL |
SAMPLE EXPERIMENTAL PROTOCOL
Culture cells to ~ 90% confluency at time of transfection.
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.
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.
References
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
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
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
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
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