Journal:

Article Title: Nuclease-Resistant Single-Stranded DNA Controls for Nucleic Acid Amplification Assays ▿

doi: 10.1128/JCM.00647-07

Figure Lengend Snippet: Electron micrographs of nrssDNA. fKN16 (A), nrssDNA-B19V-QS (B), and nrssDNA-B19V-PC (C) were expressed as described in the text, and supernatants were precipitated with polyethylene glycol-NaCl. Pellets were resuspended in low-salt buffer and prepared for electron microscopy.

Article Snippet: The digest products were separated by agarose gel electrophoresis and the B19V insert migrating at approximately 150 bp was purified from the gel (MinElute gel extraction kit; QIAGEN). fKN16 phage vector (ATCC 37002) was transformed into chemically competent E. coli K802 (ATCC 33526), and plasmid DNA was prepared from 100 ml of a K802∼fKN16 culture grown in 2xYT medium (Roth, Karlsruhe, Germany) supplemented with 20 μg of tetracycline (Roche Applied Science)/ml. fKN16 plasmid was linearized with HindIII, dephosphorylated using shrimp alkaline phosphatase (Roche Applied Science), and purified by using a High Pure PCR template purification kit (Roche Applied Science) for subsequent ligation (T4 DNA ligase; Roche Applied Science) with the HindIII-cut B19V insert.

Techniques: Electron Microscopy