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