Team:Caltech/Week 12

From 2011.igem.org

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==August 28==
==August 28==
-
Made R0010-MG1655 and J23119-MG1655 plates and overnights
+
Made R0010-MG1655 and J23119-MG1655 plates and overnights<br/>
-
Continued growing biofilm in flask with glass beads; added fresh amphicillin
+
Continued growing biofilm in flask with glass beads; added fresh amphicillin<br/>
-
Mixed more X-gal and IPTG solution at 20mg/mL and .1M concentrations respectively
+
Mixed more X-gal and IPTG solution at 20mg/mL and .1M concentrations, respectively<br/>
-
Attempted to induce R0010 overnights with IPTG to cleave X-gal; J23119 to cleave X-gal without inducing
+
Attempted to induce R0010 overnights with IPTG to cleave X-gal; J23119 to cleave X-gal without inducing<br/>
===Results===
===Results===
-
J23119-MG1655 grew, but the R0010-MG1655 did not
+
J23119-MG1655 grew, but the R0010-MG1655 did not<br/>
-
R0010 did not cleave X-gal even with a high concentration of both IPTG and X-gal
+
R0010 did not cleave X-gal even with a high concentration of both IPTG and X-gal<br/>
-
J23119 cultures turned blue with cleaved X-gal after about 4 minutes
+
J23119 cultures turned blue with cleaved X-gal after about 4 minutes<br/>
-
 
+
==August 29==
==August 29==
-
R0010 overnight culture did not grow
+
R0010 overnight culture did not grow<br/>
-
Redo p450 experiments in MeOH in order to send to electrospray
+
Redo p450 experiments in MeOH in order to send to electrospray<br/>
-
Attempt to measure J23119-X-gal reaction with spectrophotometer
+
Attempt to measure J23119-X-gal reaction with spectrophotometer<br/>
===Results===
===Results===
-
J23119 culture was too thick and not in exponential growth phase; redo
+
J23119 culture was too thick and not in exponential growth phase; redo<br/>
-
 
+
==August 30==
==August 30==
-
HPLC of yesterday's p450 reaction and purification
+
HPLC of yesterday's p450 reaction and purification<br/>
-
Overnights of DDT-pET, WT-F87A, and lacZ colonies for sequencing, BioBrick submission, spectrophotometer experiments, etc.
+
Overnights of DDT-pET, WT-F87A, and lacZ colonies for sequencing, BioBrick submission, spectrophotometer experiments, etc.<br/>
-
Make M9 media for suspending X-gal for biofilm column experiments
+
Make M9 media for suspending X-gal for biofilm column experiments<br/>
 +
Obtain pumps, tubing, and clear columns for biofilm-X-gal experiment<br/>
===Results===
===Results===
-
HPLC results revealed too much noise; chemicals at too low of concentration
+
HPLC results revealed too much noise; chemicals at too low concentration<br/>
 +
==August 31==
 +
Miniprep and sequencing of DDT-pET with T7 promoter and terminator, as well as WT-F87A using internal primers<br/>
 +
Scale up p450 degradation reaction to 500uL for HPLC<br/>
 +
===Results===
 +
R0010 glycerol stocks are not growing overnights<br/>
 +
==September 1==
 +
Sequencing of DDT-pET revealed correct results<br/>
 +
Attempt at bisdB PCR with ultrimers was not successful<br/>
 +
Made 250mL SOC media<br/>
 +
===Results===
 +
HPLC did not yield expected results<br/>
 +
==September 2==
 +
pSB3k3 vector transformed into XL10-gold strain<br/>
 +
Plate glycerol stocks of K123001 and pNT001<br/>
}}
}}

Latest revision as of 03:30, 9 September 2011


Caltech iGEM 2011



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August 28

Made R0010-MG1655 and J23119-MG1655 plates and overnights
Continued growing biofilm in flask with glass beads; added fresh amphicillin
Mixed more X-gal and IPTG solution at 20mg/mL and .1M concentrations, respectively
Attempted to induce R0010 overnights with IPTG to cleave X-gal; J23119 to cleave X-gal without inducing

Results

J23119-MG1655 grew, but the R0010-MG1655 did not
R0010 did not cleave X-gal even with a high concentration of both IPTG and X-gal
J23119 cultures turned blue with cleaved X-gal after about 4 minutes

August 29

R0010 overnight culture did not grow
Redo p450 experiments in MeOH in order to send to electrospray
Attempt to measure J23119-X-gal reaction with spectrophotometer

Results

J23119 culture was too thick and not in exponential growth phase; redo

August 30

HPLC of yesterday's p450 reaction and purification
Overnights of DDT-pET, WT-F87A, and lacZ colonies for sequencing, BioBrick submission, spectrophotometer experiments, etc.
Make M9 media for suspending X-gal for biofilm column experiments
Obtain pumps, tubing, and clear columns for biofilm-X-gal experiment

Results

HPLC results revealed too much noise; chemicals at too low concentration

August 31

Miniprep and sequencing of DDT-pET with T7 promoter and terminator, as well as WT-F87A using internal primers
Scale up p450 degradation reaction to 500uL for HPLC

Results

R0010 glycerol stocks are not growing overnights

September 1

Sequencing of DDT-pET revealed correct results
Attempt at bisdB PCR with ultrimers was not successful
Made 250mL SOC media

Results

HPLC did not yield expected results

September 2

pSB3k3 vector transformed into XL10-gold strain
Plate glycerol stocks of K123001 and pNT001


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