IGEM:MIT/2006/Notebook/2007-8-7: Difference between revisions
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=Things to Do= | =Things to Do= | ||
1. GC extract 25-mL cultures of the following that have been growing since 11 pm Sunday: | 1. GC extract 25-mL cultures of the following that have been growing since 11 pm Sunday- DONE: | ||
*J45700 EZ media -Vitamins + Fructose | *J45700 EZ media -Vitamins + Fructose OD600=2.78 | ||
*J45700 EZ media -Vitamins -TRP -TYR -PHE + Fructose | *J45700 EZ media -Vitamins -TRP -TYR -PHE + Fructose OD600=2.40 | ||
*J45800 EZ media -Vitamins + Fructose | *J45800 EZ media -Vitamins + Fructose OD600=2.30 | ||
*J45800 EZ media -Vitamins -TRP -TYR -PHE + Fructose | *J45800 EZ media -Vitamins -TRP -TYR -PHE + Fructose OD600=1.62 | ||
2. Run the four GC samples | 2. Run the four GC samples plus TOP10 cells- DONE | ||
'''Results''' | |||
4. | J45700 EZ media: 4.6 ppm, 1.65 ppm/OD600 | ||
J45700 EZ media -TRP -TYR -PHE: 4.9 ppm, 2.04 ppm/OD600 | |||
J45800 EZ media: 0.1 ppm, 0.435 ppm/OD600 | |||
7. Make fresh J45250 + J45181 cultures with control of TOP 10 cells all with precursors added and grow them up at 220 RPM | J45800 EZ media -TRP -TYR -PHE: 2.7 ppm, 1.67 ppm/OD600 | ||
3. Run the protein GFP gel samples on protein gel- DONE | |||
4. Talk with Barry/Jason about banana biosynthesis- DONE (suggested meeting with everyone there including Drew to establish whether phase-sensitive control should be focus or smell without precursor should be focus- I agree, will discuss with Drew) | |||
5. Smell J45250 + J45181 mix cultures and see if they smell- DONE (for both high copy: mint in exponential, banana in stationary; for high copy mint and mid copy banana: mint in exponential, slight mint and slight banana in stationary) | |||
6. Make fresh 25-mL J45250 + J45181 cultures (diluted 1:100) with control of TOP 10 cells all with precursors added and grow them up at 220 RPM- DONE (for some reason not growing fast (even for these cells), will let them grow over night) | |||
7. Make glycerols of J45250 + J45181 cultures from last night- DONE | |||
8. Grow up 8 mL TOP 10 to 17 mL EZ media with glycerol to GC extract as a control for today's GC experiment- DONE (grew up for ~80 mins., OD600=1.82) | |||
9. Ask Reshma if she wants to be around for the protein gel- DONE | |||
10. Make GC reservations- DONE | |||
11. Talk with Drew about setting up a meeting- DONE (2:15 tomorrow) | |||
12. Send out an email about meeting tomorrow- DONE | |||
13. Make an LC of IlvE- strain- DONE | |||
14. Check for restriction sites in tyrB gene cassette- DONE (2 PstI sites found) | |||
Links: | |||
http://www.pubmedcentral.nih.gov/picrender.fcgi?artid=1146613&blobtype=pdf | |||
http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nuccore&id=345134 | |||
15. Make 25-mL J45700 and J45800 cultures in LB to compare on the GC tomorrow with today's results- DONE | |||
Latest revision as of 16:56, 7 August 2007
Things to Do
1. GC extract 25-mL cultures of the following that have been growing since 11 pm Sunday- DONE:
- J45700 EZ media -Vitamins + Fructose OD600=2.78
- J45700 EZ media -Vitamins -TRP -TYR -PHE + Fructose OD600=2.40
- J45800 EZ media -Vitamins + Fructose OD600=2.30
- J45800 EZ media -Vitamins -TRP -TYR -PHE + Fructose OD600=1.62
2. Run the four GC samples plus TOP10 cells- DONE
Results
J45700 EZ media: 4.6 ppm, 1.65 ppm/OD600
J45700 EZ media -TRP -TYR -PHE: 4.9 ppm, 2.04 ppm/OD600
J45800 EZ media: 0.1 ppm, 0.435 ppm/OD600
J45800 EZ media -TRP -TYR -PHE: 2.7 ppm, 1.67 ppm/OD600
3. Run the protein GFP gel samples on protein gel- DONE
4. Talk with Barry/Jason about banana biosynthesis- DONE (suggested meeting with everyone there including Drew to establish whether phase-sensitive control should be focus or smell without precursor should be focus- I agree, will discuss with Drew)
5. Smell J45250 + J45181 mix cultures and see if they smell- DONE (for both high copy: mint in exponential, banana in stationary; for high copy mint and mid copy banana: mint in exponential, slight mint and slight banana in stationary)
6. Make fresh 25-mL J45250 + J45181 cultures (diluted 1:100) with control of TOP 10 cells all with precursors added and grow them up at 220 RPM- DONE (for some reason not growing fast (even for these cells), will let them grow over night)
7. Make glycerols of J45250 + J45181 cultures from last night- DONE
8. Grow up 8 mL TOP 10 to 17 mL EZ media with glycerol to GC extract as a control for today's GC experiment- DONE (grew up for ~80 mins., OD600=1.82)
9. Ask Reshma if she wants to be around for the protein gel- DONE
10. Make GC reservations- DONE
11. Talk with Drew about setting up a meeting- DONE (2:15 tomorrow)
12. Send out an email about meeting tomorrow- DONE
13. Make an LC of IlvE- strain- DONE
14. Check for restriction sites in tyrB gene cassette- DONE (2 PstI sites found)
Links:
http://www.pubmedcentral.nih.gov/picrender.fcgi?artid=1146613&blobtype=pdf
http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nuccore&id=345134
15. Make 25-mL J45700 and J45800 cultures in LB to compare on the GC tomorrow with today's results- DONE
