Navigation Links
Optimizing electroporation parameters

Other important parameters for optimizing electroporation.
Influence of the DNA/RNA The transfection efficiency of electroporation can be affected by the concentration, the purity and the size of the molecules used.

a) Influence of nucleic acid concentration:
With the optimal electroporation parameters (osmolarity, voltage, pulse length), the quality of the results obtained with plasmid concentrations between 5 g/ml and 20 g/ml is usually satisfactory. The efficiency of the transfection may be raised by increasing the nucleic acid concentration, but only within a limited concentration range. Tests with various different cell lines have shown that only in very few cases (e.g. when large plasmids were used) do plasmid concentrations in excess of 20 g/ml lead to an increase in the transfection rate (see Fig. 1.).

Figure 1. Transient transfection efficiency in NIH-3T3 cells in relation to the DNA concentration (g/ml). The cells were electroporated with different concentrations of the pEGFP-N1 plasmid. The transfection rate (max. = 100%) was determined by FACS analysis. b) Influence of nucleic acid purity:
Empirical studies have shown that EDTA and buffer salts such as HEPES or TRIS can drastically reduce transfection efficiency. Therefore, it is recommended to dissolve the nucleic acid in distilled water before transfection. Any losses resulting from DNA buffer exchange are usually more than compensated by the increased transfection efficiency. Irrespective of the preparation method used, the DNA/RNA should be ultra-pure (A260 nm/A280 nm between 1.7 and 1.9). It is also important to use endotoxin-free DNA for sensitive cells. Otherwise an increase in the DNA concentration will lead to an increase in the endotoxin content in the cell suspension.

c) Influence of the size of the plasmid:
Transfection efficiency is also affected by the size of the individual molecule that is introduced into the cell. This means that the optimal electroporation parameters that were determined for a certain plasmid, for example, have to be changed when a larger or smaller plasmid is used.



Page: All 1 2

Related biology technology :

1. Optimizing Transfection Conditions for Studying Signal Transduction Pathways
2. Optimizing pfuturbo DNA Polymerase Amplification Reactions with Perfect Match PCR Enhancer
3. Optimizing electroporation parameters
4. Optimizing Büchi® Rotary Evaporator Applications
5. Optimizing electroporation parameters
6. Optimizing Büchi® Rotary Evaporator Applications
7. Optimizing Bchi Rotary Evaporator Applications
8. Optimizing electroporation parameters
9. Optimizing electroporation parameters
10. Optimizing electroporation parameters
11. Optimizing electroporation parameters
Post Your Comments:

(Date:6/23/2016)... , June 23, 2016 A person commits a ... crime scene to track the criminal down. An ... Food and Drug Administration (FDA) uses DNA evidence to track ... Sound far-fetched? It,s not. The FDA has increasingly ... support investigations of foodborne illnesses. Put as simply as possible, ...
(Date:6/23/2016)... 2016  The Biodesign Challenge (BDC), a university competition ... harness living systems and biotechnology, announced its winning teams ... New York City . The ... projects at MoMA,s Celeste Bartos Theater during the daylong ... senior curator of architecture and design, and Suzanne ...
(Date:6/23/2016)... ... ... In a new case report published today in STEM CELLS Translational Medicine, doctors ... being treated for breast cancer benefitted from an injection of stem cells derived from ... frequent side effect of cancer treatment. , Lymphedema refers to the swelling ...
(Date:6/23/2016)... NEW YORK , June 23, 2016 ... the trading session at 4,833.32, down 0.22%; the Dow Jones ... the S&P 500 closed at 2,085.45, down 0.17%. has ... INFI ), Nektar Therapeutics (NASDAQ: NKTR ... BIND Therapeutics Inc. (NASDAQ: BIND ). Learn more ...
Breaking Biology Technology:
(Date:5/9/2016)... Elevay is currently known as ... for high net worth professionals seeking travel for work ... world, there is still no substitute for a face-to-face ... your deal with a firm handshake. This is why ... of citizenship via investment programs like those offered by ...
(Date:4/28/2016)... GOTHENBURG, Sweden , April 28, 2016 ... 1,491.2 M (139.9), up 966% compared with the first quarter of ... Operating profit totaled SEK 589.1 M (loss: 18.8) and the operating ... SEK 7.12 (loss: 0.32) Cash flow from operations was ... , The 2016 revenue guidance is unchanged, SEK 7,000-8,500 M. ...
(Date:4/19/2016)... , UAE, April 20, 2016 ... be implemented as a compact web-based "all-in-one" system solution ... the biometric fingerprint reader or the door interface with ... of modern access control systems. The minimal dimensions of ... ID readers into the building installations offer considerable freedom ...
Breaking Biology News(10 mins):