stable dna polymerase  (New England Biolabs)

Journal: Nucleic Acids Research

Article Title: Synthesizing supercoiled circular DNA molecules in vitro

doi: 10.1093/nar/gkaf889

Figure Lengend Snippet: Novel strategies to synthesize Sc double-stranded circular DNA molecules. Symbols: PCR, polymerase chain reaction; DNAP, DNA polymerase; RCA, rolling circle amplification; Rx, relaxed; Sc, supercoiled; Cre, Cre DNA recombinase; T5E, T5 exonuclease; EB, ethidium bromide; vTop1, variola virus DNA topoisomerase 1; Phenol, phenol extraction; dsDNA, double-stranded DNA. DpnI can be used to remove DNA templates derived from E. coli if needed.

Article Snippet: Thermal stable Taq DNA polymerase and SYBR Green were purchased from Fisher Scientific, Inc.

Techniques: Polymerase Chain Reaction, Amplification, Virus, Extraction, Derivative Assay

Journal: Nucleic Acids Research

Article Title: Synthesizing supercoiled circular DNA molecules in vitro

doi: 10.1093/nar/gkaf889

Figure Lengend Snippet: In vitro synthesizing Sc circular DNA pLoxFL-A using the RCA-based biochemical method. ( A ) An experimental procedure to generate Rx and Sc plasmid pLoxFL-A using the RCA-based biochemical method. ( B ) RCA of Nk pLoxFL by Φ29 DNA polymerase as described under the “Materials and methods” section. Lane 1, λ-DNA HindIII digest; Lane 2, Sc pLoxFL; Lane 3, Nk pLoxFL; Lane 4, RCA product; Lane 5, the BamHI digest of the RCA product. ( C ) Generation of Rx and Sc pLoxFL-A using the RCA-based biochemical method. RCA reactions and Cre DNA recombination were performed as described under the “Materials and methods” section. Lane 6, RCA product BamHI digest; Lanes 7–10, recombination products by Cre DNA recombinase; Lane 7, no T5 exonuclease; Lanes 8–10, 200 nM of T5 exonuclease was added; Lane 9, purified and concentrated Rx pLoxFL-A; Lane 10, Sc pLoxFL-A by E. coli DNA gyrase.

Article Snippet: Thermal stable Taq DNA polymerase and SYBR Green were purchased from Fisher Scientific, Inc.

Techniques: In Vitro, Plasmid Preparation, Purification