IGEM:MIT/2006/Notebook/2006-6-20

To do

 * 1) Possibly reconsider BAMT/SAMT primers
 * 2) Reconsider ATF1 primers definitely
 * 3) Get in touch with GC lab
 * 4) ****3 pm: Stephen has meeting --- light sensor part!

WINTERGREEN!!!!!!! (and plating)

 * 1) The liquid cultures from yesterday SMELLED LIKE WINTERGREEN!! SO EXCITING!!!! YAY!
 * 2) The SAMT+SA and the BSMT+SA worked, in both LB and minimal media
 * 3) *Now it's time to optimize SA concentration :)
 * 4) Plated liquid cultures onto M9 with 50, 100, 200 &mu;L SA
 * 5) *2 sets of 4 (BSMT M9, BSMT LB, SAMT M9, SAMT LB) with one set of 3 missing BSMT M9

Indole

 * 1) Indole from 6-19-2006 definitely was root of bad bacterial smell.

Repeat Smell Experiment
Each tube has 20 uL cells + 10 mL LB Kan + Salicylic Acid unless otherwise noted

Controls

BL21 + LB (no Kan) + 6.2 uL SA + Induced

SAMT + 31.0 uL SA

SAMT + 6.2 uL SA

SAMT + 3.1 uL SA

BSMT + 31.0 uL SA

BSMT + 6.2 uL SA

BSMT + 3.1 uL SA

Experimental

SAMT + 31.0 uL SA + Induced

3 Tubes X (SAMT + 6.2 uL SA + Induced

SAMT + 3.1 uL SA + Induced

BSMT + 31.0 uL SA + Induced

3 Tubes X (BSMT + 6.2 uL BSMT + Induced)

BSMT + 3.1 uL SA + Induced

Repeat BAMT/SAMT PCR and run PCR products on gels

 * 1) 16 sample tubes, 6 controls (8, 3 of each)
 * 2) *(-): no template- just primers, (-): no primer- just plasmid, (+): new primer/template
 * 3) Sample master mix 18x, control master mix 10x, from here
 * 4) Temperature gradient from 42-54
 * 5) PCR was a failure, must repeat at higher temperature

Repeat ATF1 PCR at higher temperature range

 * 1) try 48 to 64 degrees
 * 2) place most of tubes in center of PCR machine

BSMT PCR cleanup and digest

 * 1) cut with ECOR1 and PST1