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Measuring Oligonucleotide Hydrolysis in Cellular Lysates via Viscosity Measurements.
Menacho-Melgar, Romel; Lynch, Michael D.
Afiliação
  • Menacho-Melgar R; Department of Biomedical Engineering, Duke University, Durham, NC, USA.
  • Lynch MD; Department of Biomedical Engineering, Duke University, Durham, NC, USA.
Bio Protoc ; 12(2): e4304, 2022 Jan 20.
Article em En | MEDLINE | ID: mdl-35127994
Cell lysis, a process that releases host oligonucleotides, is required in many biotechnological applications. However, intact oligonucleotides in crude cellular lysates increase the viscosity of lysates, which complicates downstream processes and routine laboratory workflows. To address this, nucleases that hydrolyze the intact oligonucleotides are commonly added, either as purified enzymes or co-expressed in genetically engineered bacterial strains. To measure oligonucleotide hydrolysis, common DNA quantification methods, such as qPCR or fluorescence-based, require expensive reagents and equipment, and cannot distinguish different-sized DNA fragments. Here, we outline a simple alternative method for measuring DNA/RNA hydrolysis in cellular lysates, by measuring their viscosity. This method only requires common laboratory supplies and a cell phone camera.
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Texto completo: 1 Base de dados: MEDLINE Idioma: En Ano de publicação: 2022 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Idioma: En Ano de publicação: 2022 Tipo de documento: Article