IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-7-10: Difference between revisions
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*In the future, we should organize our presentations a little better and give a high-level overview at the start. | *In the future, we should organize our presentations a little better and give a high-level overview at the start. | ||
==New | ==New cultures of PCC 7942== | ||
[[Image:A new hope revisited.jpg|thumb|right|Our new culture grew quickly]] | [[Image:A new hope revisited.jpg|thumb|right|Our new culture grew quickly]] | ||
Both flasks from Friday showed growth. The flask with thiosulfate was rich green while the flask without was paler. We were surprised and pleased to find that our cyanobacteria was growing so quickly. | Both flasks from Friday showed growth. The flask with thiosulfate was rich green while the flask without was paler. We were surprised and pleased to find that our cyanobacteria was growing so quickly. |
Revision as of 11:29, 10 July 2006
Newer entries at the bottom.
Oscillation model
Jeff finished adding sinusoidal and feedback alpha functions to his model. He will upload the code later.
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.
- Also, we should do more research on PCR protocols for cyanobacteria.
- In the future, we should organize our presentations a little better and give a high-level overview at the start.
New cultures of PCC 7942
Both flasks from Friday showed growth. The flask with thiosulfate was rich green while the flask without was paler. We were surprised and pleased to find that our cyanobacteria was growing so quickly.
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.
Frozen stocks
Peng made frozen stocks of our newly grown PCC 7942. (Peng fill this in)
Mega PCR Debug
SUBSTRATE | PRIMERS | TAQ + ATP | [Mg+] | NTP | Annealing Temp | PCR Machine |
Cy7942 | Cy primers | Old Taq | - | Old NTPs | 65\' | 1 |
Cy7942 | Cy primers | New Taq | 2mM | Old NTPs | 65\' | 1 |
Cy7942 | Cy primers | Old Taq | - | New NTPs | 65\' | 1 |
Cy7942 | Cy primers | New Taq | 2mM | New NTPs | 65\' | 1 |
Cy7942 | Cy primers | New Taq | 4mM | New NTPs | 65\' | 1 |
Cy7942 | Cy primers | New Taq | 6mM | New NTPs | 65\' | 1 |
Biobrick Miniprep | Biobrick Primer | Old Taq | - | Old NTPs | 65\' | 1 |
Biobrick Miniprep | Biobrick Primer | New Taq | 2mM | Old NTPs | 65\' | 1 |
Biobrick Miniprep | Biobrick Primer | Old Taq | - | New NTPs | 65\' | 1 |
Biobrick Miniprep | Biobrick Primer | New Taq | 2mM | New NTPs | 65\' | 1 |
Biobrick Miniprep | Biobrick Primer | New Taq | 4mM | New NTPs | 65\' | 1 |
Biobrick Miniprep | Biobrick Primer | New Taq | 6mM | New NTPs | 65\' | 1 |
Cy7942 | Cy primers | New Taq | 2mM | New NTPs | 55\' | 1 |
Cy7942 | Cy primers | New Taq | 4mM | New NTPs | 55\' | 1 |
Cy7942 | Cy primers | New Taq | 6mM | New NTPs | 55\' | 1 |
Biobrick Miniprep | Biobrick Primer | New Taq | 2mM | New NTPs | 55\' | 1 |
Cy7942 | Cy primers | New Taq | 2mM | New NTPs | 65\' | 2 |
Cy7942 | Cy primers | New Taq | 4mM | New NTPs | 65\' | 2 |
Cy7942 | Cy primers | New Taq | 6mM | New NTPs | 65\' | 2 |
Biobrick Miniprep | Biobrick Primer | New Taq | 2mM | New NTPs | 65\' | 2 |
Cy7942 | Cy primers | New Taq | 2mM | New NTPs | 55\' | 2 |
Cy7942 | Cy primers | New Taq | 4mM | New NTPs | 55\' | 2 |
Cy7942 | Cy primers | New Taq | 6mM | New NTPs | 55\' | 2 |
Biobrick Miniprep | Biobrick Primer | New Taq | 2mM | New NTPs | 55\' | 2 |