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Eap binding to DNA is affected by the state and type of DNA. Eap (0.5 μg/ml) was co-incubated with DNA samples (100 ng/ml) obtained from different sources, and subsequently attached to OTS silicon prior to AFM imaging. Representative AFM height images of Eap/DNA complexes adhering to OTS silicon from three independent experiments are shown: (A) Phage lambda DNA. (B) Artificial 1.4 kb PCR DNA-product. (C) Pst I-digested plasmid pBR322 (D) Circular pBR322 isolated from E. coli DH5α. (E) Sheared herring sperm DNA. Eap molecules putatively adhering to DNA (white arrows), OTS (yellow arrows), or cross-linking DNA (green arrows) are indicated. DNA molecules displayed in (A–C) , and (E) were aligned to OTS silicon by DNA combing.

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: The Staphylococcus aureus Extracellular Adherence Protein Eap Is a DNA Binding Protein Capable of Blocking Neutrophil Extracellular Trap Formation

doi: 10.3389/fcimb.2018.00235

Figure Lengend Snippet: Eap binding to DNA is affected by the state and type of DNA. Eap (0.5 μg/ml) was co-incubated with DNA samples (100 ng/ml) obtained from different sources, and subsequently attached to OTS silicon prior to AFM imaging. Representative AFM height images of Eap/DNA complexes adhering to OTS silicon from three independent experiments are shown: (A) Phage lambda DNA. (B) Artificial 1.4 kb PCR DNA-product. (C) Pst I-digested plasmid pBR322 (D) Circular pBR322 isolated from E. coli DH5α. (E) Sheared herring sperm DNA. Eap molecules putatively adhering to DNA (white arrows), OTS (yellow arrows), or cross-linking DNA (green arrows) are indicated. DNA molecules displayed in (A–C) , and (E) were aligned to OTS silicon by DNA combing.

Article Snippet: Herring sperm DNA (10164142), phage lambda DNA (D3779), and pBR322 (N3033L) DNA were purchased from Invitrogen (Karlsruhe, Germany), Sigma-Aldrich, and New England Biolabs (Frankfurt, Germany), respectively.

Techniques: Binding Assay, Incubation, Imaging, Lambda DNA Preparation, Plasmid Preparation, Isolation