To confirm and analyze the clonal relatedness among the triclosan-resistant isolates, PFGE was performed in accordance with the PulseNet protocols published by the US Centers for Disease Control and Prevention (CDC) with some minor modifications. Briefly, the cell suspensions were treated with protease K and incubated with the XbaI restriction enzyme for at least 2 h at 37 °C to digest the DNA fragments. Then, PFGE was performed using the CHEF-MAPPER XA PFG system (Bio-Rad, USA) for 18 h. The detailed running condition were as follows: initial switch time value of 2.16 s and a final switch time of 54.17 s at a gradient of 6 V/cm at a 120° included angle [24 (link)]. Next, the electrophoretic banding patterns were visualized by the GelDoc XR gel imaging system (Bio-Rad, USA) and further analyzed by Quantity One (Bio-Rad Laboratories, USA). The Unweighted Pair Group Method with Arithmatic Mean (UPGMA) with optimization set at 1.5% to create the dendrogram at the cut-off line ≥85% was considered to analyze the genetic relatedness [25 (link)]. The standard Salmonella strain H9812 was considered as the positive control.
Chef mapper xa pfg system
Manufactured by Bio-Rad
Sourced in United States
The CHEF-MAPPER XA PFG system is a laboratory instrument designed for pulsed-field gel electrophoresis (PFGE), a technique used for the separation and analysis of large DNA molecules. The system provides a controlled and programmable environment for the electrophoretic separation of DNA fragments, enabling researchers to study genome organization, chromosomal structure, and genetic variations.
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Lab products found in correlation
2 protocols using chef mapper xa pfg system
1
Pulsed-field gel electrophoresis for triclosan-resistant isolates
2
Clonal Analysis of Triclosan-Resistant Isolates
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To con rm and analyze the clonal relatedness of resistant isolates, PFGE was also used for analysis the clonal relatedness of the triclosan resistant isolates, according to the PulseNet protocols published by the US Centers for Disease Control and Prevention (CDC) with minor modi cations. The cell suspensions treated with protease K were incubated with XbaI restriction enzyme at least for 2 hours at 37 ºC to digest the DNA fragments. Then PFGE was performed using a CHEF-MAPPER XA PFG system (Bio-Rad, USA) for 18 hours. The detailed running condition were as follows: initial switch time value of 2.16 sec, nal switch time of 54.17 sec at a gradient of 6 V/cm at a 120°i ncluded angle [23] . Next, the electrophoretic banding patterns were visualized by GelDoc XR gel imaging system (Bio-Rad, USA) and further analyzed by Quantity One (Bio-Rad Laboratories, USA). The Unweighted Pair Group Method with Arithmatic Mean (UPGMA) with optimization set at 1.5% to create the dendrogram, cut off line ≥ 85% was considered to analyze genetic relatedness [24] . Salmonella standard strain H9812 was taken as the positive control.
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