2003 | OriginalPaper | Buchkapitel
A PCR-based Protocol for In Vitro Selection of Non-crosshybridizing Oligonucleotides
verfasst von : Russell Deaton, Junghuei Chen, Hong Bi, Max Garzon, Harvey Rubin, David Harlan Wood
Erschienen in: DNA Computing
Verlag: Springer Berlin Heidelberg
Enthalten in: Professional Book Archive
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DNA computing often requires oligonucleotides that do not produce erroneous cross-hybridizations. By using in vitro evolution, huge libraries of non-crosshybridizing oligonucleotides might be evolved in the test tube. As a first step, a fitness function that corresponds to non-crosshybridization has to be implemented in an experimental protocol. Therefore, a modified version of PCR that selects non-crosshybridizing oligonucleotides was designed and tested. Experiments confirmed that the PCR-based protocol did amplify maximally mismatched oligonucleotides selectively over those that were more closely matched. In addition, a reaction temperature window was identified in which discrimination between matched and mismatched might be obtained. These results are a first step toward practical manufacture of very large libraries of non-crosshybridizing oligonucleotides in the test tube.