Electrophoresis Conditions
Escherichia coli O157:H7 and Shigella sonnei strains restricted with XbaI or AvrII (BlnI):
Select following conditions on CHEF Mapper
o Auto Algorithm
o 30 kb: low MW
o 600 kb: high MW
o Select default values except where noted by pressing "enter".
o Change run time to 18-19 hours (See note below)
o (Default values: Initial switch time = 2.16 s; Final switch time = 54.17 s)
Select following conditions on CHEF-DR III
o Initial switch time: 2.2 s
o Final switch time: 54.2 s
o Voltage: 6 V
o Included Angle: 120°
o Run time: 18-19 hours (See note below)
Select following conditions on CHEF-DR II
o Initial A time: 2.2 s
o Final A time: 54.2 s
o Start ratio: 1.0 (if applicable)
o Voltage: 200 V
o Run time: 19-20 hours (See note below)
Salmonella strains restricted with XbaI or AvrII (BlnI):
Select following conditions on CHEF Mapper
o Auto Algorithm
o 30 kb: low MW
o 700 kb: high MW
o Select default values except where noted by pressing "Enter."
o Change run time to 18-19 hours (See note below)
o (Default values: Initial switch time = 2.16 s; Final switch time = 63.8 s)
Select following conditions on CHEF DR-III
o Initial switch time: 2.2 s
o Final switch time: 63.8 s
o Voltage: 6 V
o Included Angle: 120°
o Run time: 18-19 hours (See note below)
Select following conditions on CHEF DR-II.
o Initial A time: 2.2s
o Final A time: 63.8 s
o Start Ratio: 1.0 (if applicable)
o Voltage: 200 V
o Run time: 19-20 hours (See note below)
o
PNL05 Last Updated April 2013 Page 10
Non-O157 Escherichia coli strains restricted with XbaI or AvrII (BlnI):
Select following conditions on CHEF Mapper
o Auto Algorithm
o 50 kb: low MW
o 400 kb: high MW
o Select default values except where noted by pressing "Enter."
o Change run time to 18-19 hours (See note below)
o (Default values: Initial switch time = 6.76 s; Final switch time = 35.38 s)
Select following conditions on CHEF DR-III
o Initial switch time: 6.76 s
o Final switch time: 35.38 s
o Voltage: 6 V
o Included Angle: 120°
o Run time: 18-19 hours (See note below)
Select following conditions on CHEF DR-II.
o Initial A time: 6.76 s
o Final A time: 35.38 s
o Start Ratio: 1.0 (if applicable)
o Voltage: 200 V
o Run time: 19-20 hours(See note below)
Shigella flexneri strains restricted with NotI or XBAI:
Select following conditions on CHEF Mapper
o Auto Algorithm
o 50 kb: low MW
o 400 kb: high MW
o Select default values except where noted by pressing "Enter."
o Change the switch times to the following values:
o Initial switch time: 5 seconds
o Final switch time: 35 seconds
o Change run time to 18-19 hours (See note below)
o (Default values: Initial switch time = 6.76 s; Final switch time = 35.38 s)
Select following conditions on CHEF DR-III
o Initial switch time: 5 s
o Final switch time: 35 s
o Voltage: 6 V
o Included Angle: 120°
o Run time: 18-19 hours(See note below)
Select following conditions on CHEF DR-II.
o Initial A time: 5 s
o Final A time: 35 s
o Start Ratio: 1.0 (if applicable)
o Voltage: 200 V
o Run time: 19-20 hours(See note below)
PNL05 Last Updated April 2013 Page 11
The electrophoresis running times recommended abov