IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-7-10: Difference between revisions
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We had our weekly team meeting today. Several people made some good points and suggestions: | We had our weekly team meeting today. Several people made some good points and suggestions: | ||
*For [[IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-6-30#Proposed_solution_.232|solution #2]], we can simply wash the exogenous chemical away (e.g. lactose) to disable expression at will. Pam expressed reservations about the heat shock in [[IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-6-30#Proposed_solution_.231|solution #1]]. We can substitute solution #2 for solution #1 by washing. In other words, we can fulfill both experiments of interest-- constant ''KaiABC'' production and initial ''KaiABC'' production-- with just one promoter. | *For [[IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-6-30#Proposed_solution_.232|solution #2]], we can simply wash the exogenous chemical away (e.g. lactose) to disable expression at will. Pam expressed reservations about the heat shock in [[IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-6-30#Proposed_solution_.231|solution #1]]. We can substitute solution #2 for solution #1 by washing. In other words, we can fulfill both experiments of interest-- constant ''KaiABC'' production and initial ''KaiABC'' production-- with just one promoter. | ||
*As Nick pointed out in an earlier email, even if we can't see oscillation, we can still at least show that ''something'' is happening. KaiC stays unphosphorylated in the absence of KaiA and KaiB. If we notice that KaiC is, say, 50% phosphorylated, then even if we can't observe oscillation, we know KaiABC are interacting. | *As Nick pointed out in an earlier email, even if we can't see oscillation, we can still at least show that ''something'' is happening. KaiC stays unphosphorylated in the absence of KaiA and KaiB. If we notice that KaiC is, say, 50% phosphorylated, then, even if we can't observe oscillation, we know KaiABC are interacting. | ||
*Pam suggested that we synthesize our KaiABC BB constructs in parallel with PCRing out of PCC 7942, to cover our bases in case the PCR takes a long time. | |||
*Regarding PCR, William pointed out that we might obtain a higher yield by lysing our cells chemically instead of relying on temperature. | |||
==DNA Synthesis companies== | ==DNA Synthesis companies== | ||
Revision as of 10:50, 10 July 2006
Newer entries at the bottom.
Meeting
We had our weekly team meeting today. Several people made some good points and suggestions:
- For solution #2, we can simply wash the exogenous chemical away (e.g. lactose) to disable expression at will. Pam expressed reservations about the heat shock in solution #1. We can substitute solution #2 for solution #1 by washing. In other words, we can fulfill both experiments of interest-- constant KaiABC production and initial KaiABC production-- with just one promoter.
- As Nick pointed out in an earlier email, even if we can't see oscillation, we can still at least show that something is happening. KaiC stays unphosphorylated in the absence of KaiA and KaiB. If we notice that KaiC is, say, 50% phosphorylated, then, even if we can't observe oscillation, we know KaiABC are interacting.
- Pam suggested that we synthesize our KaiABC BB constructs in parallel with PCRing out of PCC 7942, to cover our bases in case the PCR takes a long time.
- Regarding PCR, William pointed out that we might obtain a higher yield by lysing our cells chemically instead of relying on temperature.
DNA Synthesis companies
- Blue Heron Biotechnology, Inc.
- Advertised as 2-4 weeks, but teams have had problems getting sequences on time in the past.
- Codon Devices, Inc.
- DNA 2.0, Inc.
Add more links here. Additional information would be useful.
