Pulsed-field Gel Electrophoresis via Contour-clamped Homogeneous Electric Field Gels

2006 ◽  
Vol 2006 (1) ◽  
pp. pdb.prot4033
Author(s):  
Joseph Sambrook ◽  
David W. Russell
Keyword(s):  
Electric Field ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Homogeneous Electric Field ◽  
Pulsed Field

Rapid Protocol for Characterizing Klebsiella pneumoniae Isolates by Pulsed Field Gel Electrophoresis (PFGE) in Contour Clamped Homogeneous Electric Field (CHEF) Minigels

2020 ◽  
pp. 1-11
Author(s):  
Maribel Santiago Rodríguez ◽  
Juan C. Bravata Alcántara ◽  
Juan C. Martínez Briseño ◽  
Eduardo Díaz Escamilla ◽  
Ileana A. Cortes Ortiz ◽  
...  
Keyword(s):  
Klebsiella Pneumoniae ◽  
Electric Field ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Homogeneous Electric Field ◽  
Pulsed Field

Development and application of a new pulsed-field gel-electrophoresis process based on molecular dynamics

1990 ◽  
Vol 68 (9) ◽  
pp. 1055-1070 ◽  
Author(s):  
Jaan Noolandi
Keyword(s):  
Electric Field ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Biological Molecules ◽  
Dna Molecules ◽  
Theoretical Concepts ◽  
Pulsed Field ◽  
Separation Pattern ◽  
Relaxation Modes ◽  
Preparation Techniques

Separation of DNA molecules according to size is an important step in molecular biology. Pulsed-field gel electrophoresis is the main experimental tool for carrying out this separation, and is one of the techniques used in the human genome megaproject. We have developed a new approach to the separation of biological molecules by choosing electric field pulses to activate specific stretching and relaxation modes of charged molecules moving through a gel. In our process, a particular sequence of electric field pulses provides the "code" (specific instructions) for the migration of a DNA fragment of a given size to a designated position on the gel. The entire pulse train (for the duration of the experiment) can then be used to predetermine the separation pattern of a large number of DNA molecules of different sizes. This allows, for example, for the movement of only nucleic acid fragments below a certain size through the gel, and simplifies DNA preparation techniques. We have developed new hardware and software to carry out this process in a routine fashion in a laboratory environment. In this paper we review the theoretical concepts that form the basis of this separation technique, and discuss its applicability to human chromosomes and proteins.

scholarly journals Use of Pulsed-Field Gel Electrophoresis to Determine the Source of Methicillin-Resistant Staphylococcus aureus Bacteremia

2021 ◽  
Vol 13 (3) ◽  
pp. 602-610
Author(s):  
Eugene Y. H. Yeung ◽  
Ivan Gorn
Keyword(s):  
Staphylococcus Aureus ◽  
Gel Electrophoresis ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Epidemiological Studies ◽  
Pelvic Trauma ◽  
Pulsed Field Gel Electrophoresis ◽  
Bacterial Strains ◽  
Methicillin Resistant ◽  
Pulsed Field ◽  
Clinical Cases

Pulsed-field gel electrophoresis (PFGE) has historically been considered the gold standard in fingerprinting bacterial strains in epidemiological studies and outbreak investigations; little is known regarding its use in individual clinical cases. The current study detailed two clinical cases in which PFGE helped to determine the source of their methicillin-resistant Staphylococcus aureus (MRSA) bacteremia. Patient A was found to have MRSA bacteremia after trauma in her pelvic area. MRSA was also found in her groin but not in her nostril and rectum. PFGE was performed that showed variable bands of her MRSA isolates from blood and groin, suggestive of different strains of MRSA. Her MRSA bacteremia was determined to be unrelated to her pelvic trauma. Patient B was found to have MRSA bacteremia after colonoscopy. MRSA was also found in his nostril and rectum. PFGE was performed that showed variable bands of his MRSA isolates from blood and rectum but identical bands of MRSA isolates from his blood and nostril. His MRSA bacteremia was determined to be unrelated to his colonoscopy procedure. The current study demonstrates the use of PFGE to rule out the source of bacteremia in individual clinical cases.

scholarly journals Identification and Characterization of Pathogenic Yersinia enterocolitica Isolates by PCR and Pulsed-Field Gel Electrophoresis

2005 ◽  
Vol 71 (7) ◽  
pp. 3674-3681 ◽  
Author(s):  
S. Thisted Lambertz ◽  
M.-L. Danielsson-Tham
Keyword(s):  
Multiplex Pcr ◽  
Yersinia Enterocolitica ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Virulence Plasmid ◽  
Pork Meat ◽  
Pulsed Field ◽  
Pork Products ◽  
Food Borne ◽  
Pork Samples

ABSTRACT Approximately 550 to 600 yersiniosis patients are reported annually in Sweden. Although pigs are thought to be the main reservoir of food-borne pathogenic Yersinia enterocolitica, the role of pork meat as a vehicle for transmission to humans is still unclear. Pork meat collected from refrigerators and local shops frequented by yersiniosis patients (n = 48) were examined for the presence of pathogenic Yersinia spp. A combined culture and PCR method was used for detection, and a multiplex PCR was developed and evaluated as a tool for efficient identification of pathogenic food and patient isolates. The results obtained with the multiplex PCR were compared to phenotypic test results and confirmed by pulsed-field gel electrophoresis (PFGE). In all, 118 pork products (91 raw and 27 ready-to-eat) were collected. Pathogenic Yersinia spp. were detected by PCR in 10% (9 of 91) of the raw pork samples (loin of pork, fillet of pork, pork chop, ham, and minced meat) but in none of the ready-to-eat products. Isolates of Y. enterocolitica bioserotype 4/O:3 were recovered from six of the PCR-positive raw pork samples; all harbored the virulence plasmid. All isolates were recovered from food collected in shops and, thus, none were from the patients' home. When subjected to PFGE, the six isolates displayed four different NotI profiles. The same four NotI profiles were also present among isolates recovered from the yersiniosis patients. The application of a multiplex PCR was shown to be an efficient tool for identification of pathogenic Y. enterocolitica isolates in naturally contaminated raw pork.

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