SiteFind: A software tool for introducing a restriction site as a marker for successful site-directed mutagenesis |
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Authors: | Paul?M?Evans Email author" target="_blank">Chunming?LiuEmail author |
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Institution: | (1) Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, USA;(2) Sealy Center for Cancer Cell Biology, University of Texas Medical Branch, Galveston, USA |
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Abstract: | Background Site-directed mutagenesis is a widely-used technique for introducing mutations into a particular DNA sequence, often with
the goal of creating a point mutation in the corresponding amino acid sequence but otherwise leaving the overall sequence
undisturbed. However, this method provides no means for verifying its success other than sequencing the putative mutant construct:
This can quickly become an expensive method for screening for successful mutations. An alternative to sequencing is to simultaneously
introduce a restriction site near the point mutation in manner such that the restriction site has no effect on the translated
amino acid sequence. Thus, the novel restriction site can be used as a marker for successful mutation which can be quickly
and easily assessed. However, finding a restriction site that does not disturb the corresponding amino acid sequence is a
time-consuming task even for experienced researchers. A fast and easy to use computer program is needed for this task. |
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