STACKING GELS: A METHOD FOR MAXIMISING OUTPUT FOR PULSED-FIELD GEL ELECTROPHORESIS

Stacking gels: A method for maximising output for pulsed-field gel electrophoresis.

Pulsed field gel electrophoresis (PFGE), the gold standard of molecular typing methods, has a major disadvantage of an unusually long electrophoretic time. From the original protocol of 6 days, it was modified to 3 days and subsequently to a single day. We describe the procedure of stacking five to six gels one on top of another in order to increase and maximize the output in a shorter time wit...

Increased throughput of BAC/PAC insert size determinations by stacking gels during pulsed-field gel electrophoresis.

Using a Temperature Gradient against the Time in Polyacrylamide Gel Electrophoresis May Eliminate the Need for Stacking Gels

Background and Objectives: Making stacking gels for polyacrylamide gels in the laboratory by conventional methods is laborious and time consuming. Considering the role of temperature in polyacrylamide gels with respect to electrical resistance and viscosity, we assumed that decreasing the temperature would cause an increase in electrical resistance and viscosity.  Ultimately, a downward tempera...

Pulsed-field gel electrophoresis.

DNA molecules in an agarose matrix elongate and migrate toward the anode when exposed to an electric field. An agarose matrix represents a highly irregular network with pores of various dimensions, large open areas, and regions with different densities. During electrophoresis, migration will be retarded by obstructions that prevent sieving by size and result in "end-on" migration as if through ...

Method for optimizing pulsed-field gel electrophoresis banding pattern data.

The genomic DNA of 47 strains of TSST-1 toxin-producing Staphylococcus aureus were cleaved with SmaI restriction endonuclease and resolved in an agarose gel by pulsed-field gel electrophoresis (PFGE). An algorithm was designed to standardize the band weights or brightness (trace quantity) produced to a bounded region between 0 and 1 regardless of DNA fragment size while simultaneously reducing ...