Using the gene pulser MXcell electroporation system to transfect primary cells with high efficiency
- PMID: 20057352
- PMCID: PMC2807742
- DOI: 10.3791/1662
Using the gene pulser MXcell electroporation system to transfect primary cells with high efficiency
Abstract
It is becoming increasingly apparent that electroporation is the most effective way to introduce plasmid DNA or siRNA into primary cells. The Gene Pulser MXcell electroporation system and Gene Pulser electroporation buffer (Bio-Rad) were specifically developed to easily transfect nucleic acids into mammalian cells and difficult-to-transfect cells, such as primary and stem cells. We will demonstrate how to perform a simple experiment to quickly identify the best electroporation conditions. We will demonstrate how to run several samples through a range of electroporation conditions so that an experiment can be conducted at the same time as optimization is performed. We will also show how optimal conditions identified using 96-well electroporation plates can be used with standard electroporation cuvettes, facilitating the switch from electroporation plates to electroporation cuvettes while maintaining the same electroporation efficiency. In the video, we will also discuss some of the key factors that can lead to the success or failure of electroporation experiments.
Similar articles
-
The preparation of primary hematopoietic cell cultures from murine bone marrow for electroporation.J Vis Exp. 2009 Jan 6;(23):1026. doi: 10.3791/1026. J Vis Exp. 2009. PMID: 19229174 Free PMC article.
-
Optimizing electroporation conditions in primary and other difficult-to-transfect cells.J Biomol Tech. 2008 Dec;19(5):328-34. J Biomol Tech. 2008. PMID: 19183796 Free PMC article.
-
Substrate-mediated, high-efficiency siRNA electroporation.Methods Mol Biol. 2014;1121:139-46. doi: 10.1007/978-1-4614-9632-8_12. Methods Mol Biol. 2014. PMID: 24510819
-
Electroporation Knows No Boundaries: The Use of Electrostimulation for siRNA Delivery in Cells and Tissues.J Biomol Screen. 2015 Sep;20(8):932-42. doi: 10.1177/1087057115579638. Epub 2015 Apr 7. J Biomol Screen. 2015. PMID: 25851034 Free PMC article. Review.
-
Microscale electroporation: challenges and perspectives for clinical applications.Integr Biol (Camb). 2009 Mar;1(3):242-51. doi: 10.1039/b819201d. Epub 2009 Jan 29. Integr Biol (Camb). 2009. PMID: 20023735 Free PMC article. Review.
Cited by
-
Efficient transgenesis and homology-directed gene targeting in monolayers of primary human small intestinal and colonic epithelial stem cells.Stem Cell Reports. 2022 Jun 14;17(6):1493-1506. doi: 10.1016/j.stemcr.2022.04.005. Epub 2022 May 5. Stem Cell Reports. 2022. PMID: 35523179 Free PMC article.
-
Generation of murine induced pluripotent stem cells by using high-density distributed electrodes network.Biomicrofluidics. 2015 Sep 11;9(5):054107. doi: 10.1063/1.4930866. eCollection 2015 Sep. Biomicrofluidics. 2015. PMID: 26396661 Free PMC article.
-
Comparative Analysis of Non-viral Transfection Methods in Mouse Embryonic Fibroblast Cells.J Biomol Tech. 2017 Jul;28(2):67-74. doi: 10.7171/jbt.17-2802-003. Epub 2017 Apr 29. J Biomol Tech. 2017. PMID: 28507462 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources