Plasmid extractions were carried out by the use of GeneJET Plasmid Miniprep Kit following manufacturer indications (ThermoFisher Scientific, Waltham, Massachusetts, USA) and S1-Pulsed-Field Gel Electrophoresis (PFGE). Bacterial DNA embedded in agarose plugs was digested using 14 units of S1-nuclease (Takara Bio, Kusatsu, Japan) per plug followed by PFGE. Samples were run on a CHEF-DR III system (Bio-Rad, Munich, Germany) for 20 h at 6 V/cm and 14°C. CHEF DNA Size Standard Lambda Ladder (Bio-Rad) was used as molecular weight marker. Southern blot hybridizations were performed to locate blaNDM-1, blaPER-7 and blaGES-like genes with specific digoxigenin-labeled DNA probes (Roche, Mannheim, Germany). Signal detection was performed using DIG Nucleic Acid Detection Kit (Roche). Determination of the presence and classification of replicase genes was conducted using A. baumannii PCR-Based Replicon Typing as previously described (Bertini et al., 2010 (link)).
Chef dna size standard lambda ladder
Manufactured by Bio-Rad
Sourced in United States
The CHEF DNA Size Standard Lambda Ladder is a laboratory tool used for estimating the size of DNA fragments during gel electrophoresis. It consists of a series of DNA fragments of known sizes derived from the lambda phage genome, which serve as a reference for comparison to the DNA samples being analyzed.
Automatically generated - may contain errors
Lab products found in correlation
Genejet plasmid miniprep kit, by Thermo Fisher Scientific (1 mentions)
Dig nucleic acid detection kit, by Roche (1 mentions)
S1 nuclease, by Takara Bio (1 mentions)
Chef dr 3 system, by Bio-Rad (1 mentions)
Chef dr 2 pulsed field electrophoresis system, by Bio-Rad (1 mentions)
Gel doc camera system, by Bio-Rad (1 mentions)
2 protocols using chef dna size standard lambda ladder
1
Plasmid Extraction and Molecular Characterization
2
Pulsed-Field Gel Electrophoresis (PFGE) of Y. enterocolitica O8
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PFGE was performed to compare the genetic characteristics of Y. enterocolitica O8 isolates. The PFGE was carried out as described by Iwata et al. [13 (link)]. Briefly, chromosomal DNAs were digested by the restriction enzyme NotI (TaKaRa, Kusatsu, Japan) for 3 hr at 37°C. The DNA fragments
were separated in 1.2% agarose NA (GE Healthcare, Bioscience AB, Uppsala, Sweden) on a CHEF-DRII Pulsed Field Electrophoresis System (Bio-Rad Laboratories, Inc., Hercules, CA, USA).
Electrophoresis was carried out for 24 hr at 14°C and 200 V with pulse times of 2 to 25 sec. A CHEF DNA Size Standard Lambda Ladder (Bio-Rad) was used as molecular size marker. The gels were
stained with AtlasSight DNA Stain (Bioatlas, Tartu, Estonia) and photographed using a Gel Doc camera system (Bio-Rad).
were separated in 1.2% agarose NA (GE Healthcare, Bioscience AB, Uppsala, Sweden) on a CHEF-DRII Pulsed Field Electrophoresis System (Bio-Rad Laboratories, Inc., Hercules, CA, USA).
Electrophoresis was carried out for 24 hr at 14°C and 200 V with pulse times of 2 to 25 sec. A CHEF DNA Size Standard Lambda Ladder (Bio-Rad) was used as molecular size marker. The gels were
stained with AtlasSight DNA Stain (Bioatlas, Tartu, Estonia) and photographed using a Gel Doc camera system (Bio-Rad).
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