Bioline velocity dna polymerase

Chromosomal DNA was isolated from N. gonorrhoeae cells by the use of a Wizard genomic DNA purification kit (Promega Co., USA) and plasmid DNA from E. coli using an Isolate II plasmid minikit (Bioline, Boston, USA) per the instructions of the manufacturers. Primers used in this study are listed in Table S1 in the supplemental material and were synthesized by GeneWorks (GeneWorks Pty. Ltd., Australia) or Integrated DNA Technologies. Bioline Velocity DNA polymerase was used in all cloning and mutagenesis procedures and Bioline MangoTaq DNA polymerase for screening PCRs (Bioline, Boston, MA, USA). All restriction digestion enzymes and T4 DNA ligase were from New England Biolabs (New England Biolabs Inc., Ipswich, MA, USA). Transformations into N. gonorrhoeae or E. coli cells were carried as described previously (42 (link), 43 (link)). All sequencing was performed by Australian Genome Research Facility (AGRF; Australia).

Plasmid DNA was isolated from E. coli cells using a Bioline Isolate II plasmid minikit (Bioline Meridian Bioscience, USA). Chromosomal DNA was isolated from N. gonorrhoeae cells by the use of a Wizard genomic DNA purification kit (Promega Co., USA). Primers used in this study are listed in Table S1 and were synthesized by Integrated DNA Technologies or Sigma-Aldrich. Bioline Velocity DNA polymerase was used in all PCR mutagenesis reactions and Bioline MangoTaq DNA polymerase for screening PCRs (Bioline, Boston, MA). All restriction digestion enzymes were from New England Biolabs (New England Biolabs, Inc., Ipswich, MA). Transformations into N. gonorrhoeae or E. coli cells were carried as described previously (38 (link), 49 (link)). All sequencing was performed by the Australian Genome Research Facility.