Two-dimensional (2D) gel electrophoresis was modified from (49 (link)). A total of 5 μg genomic DNA samples, either undigested or digested with HinfI, were electrophoresed on a 0.4% agarose gel (first dimension) at room temperature and 1V/cm for 16 h. The lanes were cut (removing the first 1 cm from the wells), embedded in a second dimension gel (1.2% agarose with 0.3 μg/ml ethidium bromide) and electrophoresed at 4.5V/cm for 6 h. The gel was dried, cut to separate the duplicated samples and hybridized side by side with C- and G-probes to detect single-stranded telomeric DNA, as described for in-gel analysis. After exposure, the gels were denatured and hybridized again with the same probes to detect the total telomeric DNA. Supercoiled DNA ladder (NEB Inc), containing nine supercoiled plasmids (2–10 kb), was used as reference.
Supercoiled dna ladder
Manufactured by New England Biolabs
Sourced in United Kingdom
The Supercoiled DNA ladder is a molecular weight marker used to determine the size of supercoiled DNA samples in agarose gel electrophoresis. It consists of a mixture of supercoiled DNA fragments of known molecular weights, allowing for the identification and quantification of supercoiled DNA samples.
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3 protocols using supercoiled dna ladder
1
Telomeric DNA Detection via 2D Gel Electrophoresis
2
Evaluation of DNA Samples Using TOTO-3
3
Plasmid Profiling of Flavobacterium psychrophilum
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Plasmid profiles of 169 isolates from representative genotypes and origins were investigated. The extraction of plasmid DNA from F. psychrophilum isolates was performed as previously described (Bartie et al. 2012) . The approximate molecular size of the plasmids was estimated using a Supercoiled DNA ladder (New England BioLabs, UK) and the known plasmid contents of two reference strains Escherichia coli V517 and 39R861 (Macrina et al. 1978) . Plasmid profiles were identified by differences in size and number of resulting DNA bands.
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