Brown: Team Resistance/2 June 2008

2 June 2008

• Cell lysis tests were done using the Freeze Thaw (F/T) method of lysis. We have yet to receive the pVJ4 and other plasmids from John Mekalanos at Harvard Medical School or Ry Young at Texas A&M University. Our tests consisted of three F/T cycles (20 minutes in the -80 deg C freezer and 30-40 min of thawing in warm water). Resistance measurements taken with a standard voltmeter.

• TUBE Initial F/T Cycle 1 F/T Cycle 2 F/T Cycle 3
• DH20 1.119 M ohm .726 M ohm .934 M ohm 1.009 M ohm
• LB+kan 36.14 k ohm 21.47 k ohm 24.98 k ohm 24.31 k ohm

1. 351.0 k ohm 280.0 k ohm 354.5 k ohm 434.0 k ohm
2. 55.3 k ohm 21.41 k ohm 22.34 k ohm 23.44 k ohm

• Trial 2 Initial F/T Cycle 1 F/T Cycle 2 F/T Cycle 3
• DH20 272.1 k ohm 235.3 k ohm 249.9 k ohm 252.2 k ohm
• LB+kan 70.8 k ohm 26.97 k ohm 27.36 k ohm 25.68 k ohm

1. 340.1 k ohm 254.9 k ohm 335.0 k ohm 382.7 k ohm
2. 40.73 k ohm 25.51 k ohm 28.94 k ohm 27.16 k ohm

• Trial 3 Initial F/T Cycle 1 F/T Cycle 2 F/T Cycle 3
• DH20 .695 M ohm .726 M ohm .742 M ohm .810 M ohm
• LB+kan 36.35 k ohm 28.03 k ohm 28.54 k ohm 28.44 k ohm

1. 326.1 k ohm 246.2 k ohm 335.6 k ohm 377.1 k ohm
2. 58.19 k ohm 30.86 k ohm 32.00 k ohm 29.54 k ohm

• Plate 20, S17-1 pdkl02 – SRRz cassette under an IPTG promoter

• Tube #1: Centrifuged 9 times to ensure a very concentrated solution then resuspended in DH20 before resistance readings.
• Tube #2: Centrifuged 9X then resuspended in LB+kan before resistance readings taken.

• Plate frozen for 20 minutes and allowed to thaw for 25 minutes before taking reading