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Team:Harvard/Dailybook/Week7/Widgetry
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(New page: =Goals for this week= =Monday: August 4, 2008= =Tuesday: August 5, 2008= =Wednesday: August 6, 2008= =Thursday: August 7, 2008= =Friday: August 8, 2008=) |
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=Goals for this week= | =Goals for this week= | ||
| + | * Shewanella and E. coli co-culture experiments | ||
| + | ** If Shewanella responds rapidly to lactate, and E. coli breaks down lactose into lactate, can we couple lactose breakdown by E. coli with current production by Shewanella? | ||
| + | ** Motivation: | ||
| + | *** An inducible system in E. coli could be used to control Shewanella current production | ||
| + | |||
=Monday: August 4, 2008= | =Monday: August 4, 2008= | ||
| + | |||
| + | == Co-culture experiment setup == | ||
| + | |||
| + | * Chambers 1 & 2: wt E. coli + wt Shewie + lactose | ||
| + | * Chambers 4 &5 : Lac-operon knockout E. coli + wt Shewie + lactose | ||
| + | |||
| + | '''Positive Controls''' | ||
| + | * Chamber 3: wt E. coli + wt Shewie + lactate | ||
| + | * Chamber 6: Lac-operon knockout E. coli + wt Shewie + lactate | ||
| + | |||
| + | '''Negative Controls''' | ||
| + | * Chamber 7: wt Shewie + lactose | ||
| + | * Chamber 8: wt E. coli + lactose | ||
| + | * Chamber 9: Lac-operon knockout E. coli + lactose | ||
| + | |||
=Tuesday: August 5, 2008= | =Tuesday: August 5, 2008= | ||
| - | =Wednesday: August 6, 2008= | + | |
| - | = | + | ==Co-culture experiment results, day 1== |
| - | = | + | |
| + | :[[Image: Co-culture_part1.JPG | 700 px]] | ||
| + | :* Notes: | ||
| + | :** At t = 2000 s, all cells added | ||
| + | :** At t = 80000 s, all injection done | ||
| + | |||
| + | |||
| + | === Zoomed-in graph of results, part 1 === | ||
| + | :[[Image: Co-culture_part1_zoomed.jpg | 700 px]] | ||
| + | :* Notes: | ||
| + | :** Zoomed-in on time since injections | ||
| + | :** Positive controls omitted | ||
| + | |||
| + | |||
| + | '''Preliminary Conclusions''' | ||
| + | :* Noticeable difference in behavior between wt E. coli + Shewie and Lac-operon knockout + Shewie co-cultures | ||
| + | :* Positive and negative controls show most likely no other variables involved | ||
| + | :* Could be used to create tic-tac-toe which uses the difference in behavior to determine moves | ||
| + | |||
| + | =Wednesday: August 6, 2008 to Friday: August 8, 2008= | ||
| + | |||
| + | ==Co-culture experiment results, day 1 & 2 == | ||
| + | |||
| + | === wt E. coli vs. Lac-operon knockout E. coli === | ||
| + | : [[Image: Co-culture_part2_test.jpg | 700px]] | ||
| + | :* Notes: | ||
| + | :** At t = 260000 s, adjusted gas for Chamber 5 | ||
| + | :** At t = 340000 s, lactose injection for Chamber 1 | ||
| + | :** Initially, very noticeable difference between wt E. coli and Lac-operon knockout E. coli, but the current production of one of the E. coli chambers rises after a day, making the difference less distinguishable | ||
| + | |||
| + | === Results w/ positive controls === | ||
| + | :[[Image: Co-culture_part2_wpos.jpg | 700px]] | ||
| + | :* Notes: | ||
| + | :* At t = 340000 s, lactose injections for Chambers 1, 3, and 6 | ||
| + | :** Wanted to see if the positive controls would exhibit the same behavior when lactose injected instead of lactate | ||
| + | |||
| + | |||
| + | === Results (entire) === | ||
| + | :[[Image: Co-culture_part2.jpg | 700px]] | ||
| + | |||
| + | :'''Concerns''' | ||
| + | :* Chambers with co-cultures contained double the amount of cells of the controls | ||
| + | :* Current production in non-positive co-culture chambers | ||
| + | :** wt Shewie + lactose | ||
| + | :** lac-operon knockout E. coli + Shewie + lactose | ||
| + | :** lac-operon knockout E. coli + lactose | ||
| + | |||
| + | :'''Conclusions''' | ||
| + | :* Although behavior of wt E. coli + Shewie and Lac-operon knockout E. coli + Shewie co-cultures are not as distinguishable after a day, our focus can only be on the first day | ||
| + | :* Will try the same experiment with DH5(alpha) and DH5(alpha)/pUC19 strains | ||
| + | :* Need to look for inducible lacZ systems | ||
| + | :** Light-repressible (RU1012 with plasmid) system? | ||
Latest revision as of 21:24, 29 October 2008
Contents |
Goals for this week
- Shewanella and E. coli co-culture experiments
- If Shewanella responds rapidly to lactate, and E. coli breaks down lactose into lactate, can we couple lactose breakdown by E. coli with current production by Shewanella?
- Motivation:
- An inducible system in E. coli could be used to control Shewanella current production
Monday: August 4, 2008
Co-culture experiment setup
- Chambers 1 & 2: wt E. coli + wt Shewie + lactose
- Chambers 4 &5 : Lac-operon knockout E. coli + wt Shewie + lactose
Positive Controls
- Chamber 3: wt E. coli + wt Shewie + lactate
- Chamber 6: Lac-operon knockout E. coli + wt Shewie + lactate
Negative Controls
- Chamber 7: wt Shewie + lactose
- Chamber 8: wt E. coli + lactose
- Chamber 9: Lac-operon knockout E. coli + lactose
Tuesday: August 5, 2008
Co-culture experiment results, day 1
- Notes:
- At t = 2000 s, all cells added
- At t = 80000 s, all injection done
- Notes:
Zoomed-in graph of results, part 1
- Notes:
- Zoomed-in on time since injections
- Positive controls omitted
- Notes:
Preliminary Conclusions
- Noticeable difference in behavior between wt E. coli + Shewie and Lac-operon knockout + Shewie co-cultures
- Positive and negative controls show most likely no other variables involved
- Could be used to create tic-tac-toe which uses the difference in behavior to determine moves
Wednesday: August 6, 2008 to Friday: August 8, 2008
Co-culture experiment results, day 1 & 2
wt E. coli vs. Lac-operon knockout E. coli
-
- Notes:
- At t = 260000 s, adjusted gas for Chamber 5
- At t = 340000 s, lactose injection for Chamber 1
- Initially, very noticeable difference between wt E. coli and Lac-operon knockout E. coli, but the current production of one of the E. coli chambers rises after a day, making the difference less distinguishable
- Notes:
Results w/ positive controls
- Notes:
- At t = 340000 s, lactose injections for Chambers 1, 3, and 6
- Wanted to see if the positive controls would exhibit the same behavior when lactose injected instead of lactate
Results (entire)
- Concerns
- Chambers with co-cultures contained double the amount of cells of the controls
- Current production in non-positive co-culture chambers
- wt Shewie + lactose
- lac-operon knockout E. coli + Shewie + lactose
- lac-operon knockout E. coli + lactose
- Conclusions
- Although behavior of wt E. coli + Shewie and Lac-operon knockout E. coli + Shewie co-cultures are not as distinguishable after a day, our focus can only be on the first day
- Will try the same experiment with DH5(alpha) and DH5(alpha)/pUC19 strains
- Need to look for inducible lacZ systems
- Light-repressible (RU1012 with plasmid) system?
