The genomic DNA was digested with S1 endonuclease (New England Biolabs, Beverly, MA, USA) separated by PFGE as previously described (38 (link)). After transfer to Amersham Hybond-N+ membranes (GE Healthcare), the genomic DNA was hybridized with a poxtA probe as described in other studies (30 (link)).
S1 endonuclease
Manufactured by New England Biolabs
Sourced in United States
S1 endonuclease is a single-strand-specific nuclease that cleaves single-stranded DNA and RNA. It recognizes and cleaves at regions of low base pairing or single-stranded overhangs.
Automatically generated - may contain errors
2 protocols using s1 endonuclease
1
Detecting Antibiotic Resistance Genes using PFGE
2
Plasmid Content Analysis by PFGE
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The plasmid content from wild-type effluent strains, transformants, and transconjugants was analyzed by pulse field gel electrophoresis (PFGE) after S1 nuclease digestion (Barton et al., 1995) (link). All PFGE reagents used were from Bio-Rad (CHEF Genomic DNA Plug Kits). Genomic DNA agarose blocks were digested with S1endonuclease (New England Biolab) for 90 min at 37°C, and then loaded into 1% electrophoretic agarose gel and run with the CHEF-DR II system (Bio-Rad), with a specific program 1-10s,18h, 6V/cm (voltage, pulse time, and electrophoretic time) at 14°C. The gels were then stained with Gel Star® (Lonza, Rockland, ME USA) and analyzed by FPQuest® software (BioRad, Marne la Coquette, France).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!