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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 | + | 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/> | ||
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Latest revision as of 03:30, 9 September 2011
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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 September 2pSB3k3 vector transformed into XL10-gold strain
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