low DNA concentrations
may result in loss of precious material. Adding glycogen as a carrier
helps to reduce this problem. Ethanol precipitation is also considered
by some people to be a relatively time-consuming step in the preparation
of DNA for electroporation. I conclude that the easiest and quickest method
for achieving high electroporation efficiencies is to heat-treat the ligation
reaction, followed by a 1
/100 dilution. Only a fraction of the ligation
reactions are used for the dilutions, leaving the remaining DNA for other
1. Dower, W. J., In Genetic Engineering-Principles and Methods, Setlow,
J. K., ed, 1990, vol. 12, pp. 275 - 296, Plenum Press, New York.
2. Willson, T. A. and Gough, N. M., Nucl. Acids Res. 16, 11820 (1988).
3. Zabarovsky, E. R. and Winberg, G., Nucl. Acids Res. 18, 5912 (1990).
4. Jacobs, M., Wnendt, S. and Stahl, U., Nucl. Acids Res. 18, 1653 (1990).
5. Ymer, S., Nucl. Acids Res. 19, 6960 (1991).
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. Electrotransformation of E. coli With M13 DNA2
. Highest Possible Transformation Efficiencies for High-Throughput
. New Competent Cells for Highest Transformation Efficiencies4
. Molecular Biology:Easy Dephosphorylation and Rapid DNA Ligation5
. Cloning Based on Efficient Three-Fragment Assembly
. Structural Analysis of Glycosylated Peptides in Complex Mixtures with
Ion Trap MSn7
. LC-MALDI and MALDI-MS/MS for the Analysis of Complex Peptide Mixtures