Journal: MethodsX

Article Title: Generating single-stranded DNA circles with minimal resources

doi: 10.1016/j.mex.2021.101300

Figure Lengend Snippet: The results from ten independent rolling transcription assays with circularized DNA templates synthesized using our method (O) and original circularization method (K), with (+RNAP) and without RNA polymerase. (A) The templates O1 and K1 are treated with Exonuclease I. (B) The templates O1 and K1 are treated with Exonuclease I, while the templates O2 and K2 are treated with S1 Nuclease. Notes: The assays were conducted using previously published experimental procedure [ , ]. Each assay contained an exact same amount of DNA template based on the DNA quantification through NanoDrop spectrometric analysis. All DNA templates were isolated, quantified, and utilized in these assays without continuing through the dialysis (Steps 9 and 10 in the method).

Article Snippet: Resource availability: , Unless otherwise specified, all reagents were purchased from Sigma-Aldrich (St- Louis, MO). The 5′-phosphorylated linear DNA substrate of Kool™ NC-45™ Universal RNA Polymerase template [ ] and splint DNA substrates were purchased from Integrated DNA Technologies (see the supporting information for the specific sequences; the DNA sequence for Kool™ NC-45™ Universal RNA Polymerase template [ , ], was provided by epicentre (Madison, WI) once the company discontinued the product, Kool™ NC-45™ Universal RNA Polymerase template). T4 DNA Ligase and Exonuclease I ( E. coli ) were purchased from New England Biolabs (Ipswich, MA). S1 Nuclease ( Aspergillus oryzae ) was purchased from Promega (Madison, WI). The dialysis device (Tube-O-DIALYZER™, Medi, 1 K MWCO, 20) was purchased from G-Biosciences (St. Louis, MO). The purified wild-type E. coli RNA polymerase [ , , ] was a generous gift from George A. Garcia laboratory of University of Michigan (Ann Arbor, MI). .

Techniques: Synthesized, Isolation

Journal: MethodsX

Article Title: Generating single-stranded DNA circles with minimal resources

doi: 10.1016/j.mex.2021.101300

Figure Lengend Snippet: The results from ten independent rolling transcription assays with circularized DNA templates synthesized using our method (O) and original circularization method (K), with (+RNAP) and without RNA polymerase. (A) The templates O1 and K1 are treated with Exonuclease I. (B) The templates O1 and K1 are treated with Exonuclease I, while the templates O2 and K2 are treated with S1 Nuclease. Notes: The assays were conducted using previously published experimental procedure [ , ]. Each assay contained an exact same amount of DNA template based on the DNA quantification through NanoDrop spectrometric analysis. All DNA templates were isolated, quantified, and utilized in these assays without continuing through the dialysis (Steps 9 and 10 in the method).

Article Snippet: S1 Nuclease ( Aspergillus oryzae ) was purchased from Promega (Madison, WI).

Techniques: Synthesized, Isolation