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/> |
===Results=== | ===Results=== | ||
- | HPLC results revealed too much noise; chemicals at too low | + | 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/> | ||
+ | Made 250mL SOC media<br/> | ||
+ | ===Results=== | ||
+ | HPLC did not yield expected results<br/> | ||
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Revision as of 00:07, 2 September 2011
Project |
August 28Made R0010-MG1655 and J23119-MG1655 plates and overnights ResultsJ23119-MG1655 grew, but the R0010-MG1655 did not August 29R0010 overnight culture did not grow ResultsJ23119 culture was too thick and not in exponential growth phase; redo August 30HPLC of yesterday's p450 reaction and purification ResultsHPLC results revealed too much noise; chemicals at too low concentration August 31Miniprep and sequencing of DDT-pET with T7 promoter and terminator, as well as WT-F87A using internal primers ResultsR0010 glycerol stocks are not growing overnights September 1Sequencing of DDT-pET revealed correct results ResultsHPLC did not yield expected results
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