Multi-locus sequence typing (MLST) was performed by the amplifications of the internal fragments of seven housekeeping genes of carbapenemase-producing E. coli isolates (adk, fumC, icd, purA, gyrB, recA, and mdh) according to the database7 . The clonal relationships of the isolates harboring the blaNDM–5 gene were further determined by pulsed-field gel electrophoresis (PFGE). Briefly, genomic DNA of the blaNDM–5-positive E. coli isolates was prepared in agarose blocks and digested with restriction enzyme XbaI. DNA fragments were separated using a CHEF II D-Mapper XA PFGE system (Bio-Rad, Hercules, California, CA, United States) with running conditions as described previously (Jain et al., 2018 (link)).
Chef 2 d mapper xa pfge system
Manufactured by Bio-Rad
Sourced in United States
The CHEF II D-Mapper XA PFGE system is a laboratory instrument designed for pulsed-field gel electrophoresis (PFGE). It is used to separate and analyze large DNA molecules. The system includes a programmable power supply, cooling system, and gel casting platform to facilitate the PFGE process.
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Lab products found in correlation
2 protocols using chef 2 d mapper xa pfge system
1
Molecular Typing of Carbapenem-Resistant E. coli
2
Molecular Typing of Carbapenem-Resistant Enterobacteriaceae
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Molecular typing of CRE isolates was performed by PFGE. Genomic DNA of the CRE was prepared in agarose blocks and was digested with restriction enzyme XbaI (TaKaRa Biotechnology, Dalian, China). DNA fragments were separated using a CHEF II D-Mapper XA PFGE system (Bio-Rad, Hercules, CA) with running conditions as described previously.18 (link) MLST of CRE was conducted by PCR as previously described.19 (link) The allelic profiles and sequence types were identified by amplifying and sequencing the seven housekeeping genes (adk, fumC, gyrB, icd, mdh, purA, recA) according to the reference website (https://enterobase.warwick.ac.uk/species/index/ecoli ). The phylogenetic groups of CRE were determined by multiplex PCR analysis.20 (link)
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