Articles by Eliza L. Lewis in JoVE
Ensaio da atividade de DNA polimerase usando infravermelho fluorescente etiquetado DNA visualizado pela electroforese do Gel do acrilamido Eliza L. Lewis1, Aaron M. Leconte1 1Department of Chemistry, W.M. Keck Science Department of Claremont Mckenna, Pitzer, and Scripps College Este protocolo descreve a caracterização da síntese de DNA polimerase do ADN modificado através da observação de alterações para o infravermelho próximo fluorescente etiquetada DNA usando electroforese em gel e gel de imagem. Géis de acrilamida são utilizados para geração de imagens de alta resolução da separação de curtos ácidos nucleicos, que migram em taxas diferentes dependendo do tamanho.
Other articles by Eliza L. Lewis on PubMed
Design and Discovery of New Combinations of Mutant DNA Polymerases and Modified DNA Substrates Chembiochem : a European Journal of Chemical Biology. Apr, 2017 | Pubmed ID: 28160372 Chemical modifications can enhance the properties of DNA by imparting nuclease resistance and generating more-diverse physical structures. However, native DNA polymerases generally cannot synthesize significant lengths of DNA with modified nucleotide triphosphates. Previous efforts have identified a mutant of DNA polymerase I from Thermus aquaticus DNA (SFM19) as capable of synthesizing a range of short, 2'-modified DNAs; however, it is limited in the length of the products it can synthesize. Here, we rationally designed and characterized ten mutants of SFM19. From this, we identified enzymes with substantially improved activity for the synthesis of 2'F-, 2'OH-, 2'OMe-, and 3'OMe-modified DNA as well as for reverse transcription of 2'OMe DNA. We also evaluated mutant DNA polymerases previously only tested for synthesis for 2'OMe DNA and showed that they are capable of an expanded range of modified DNA synthesis. This work significantly expands the known combinations of modified DNA and Taq DNA polymerase mutants.