IGEM:MIT/2006/Notebook/2007-8-22: Difference between revisions
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*We want 200 ppm internal and external standard. | *We want 200 ppm internal and external standard. | ||
*For hexyl acetate, .87 g/mL/.0001 g/mL= 1:8700 dilution; in | *For hexyl acetate, .87 g/mL/.0001 g/mL= 1:8700 dilution; in 20 mL, 20 mL/8700 = 2.3 uL; add 2.3 uL to cultures before extraction | ||
*For oxyl acetate, .856 g/mL/.0002 g/mL= 1:8560 dilution; in 1 mL, 1 mL/8560 = .1168 uL; Make a 1:10 dilution in heptane and add 1.168 uL to 1 mL sample | *For oxyl acetate, .856 g/mL/.0002 g/mL= 1:8560 dilution; in 1 mL, 1 mL/8560 = .1168 uL; Make a 1:10 dilution in heptane and add 1.168 uL to 1 mL sample | ||
Revision as of 11:06, 22 August 2007
Schedule
1. Run GC samples of lysed cells- DONE (NOTHING SHOWED UP! NO!)
2. 10:55- J45120 15 hour sample extraction OD=2.06, 11:15- J45181 15 hour sample extraction OD=1.98, 11:55- J45120 16 hour sample extraction OD=2.18, 12:15- J45181 16 hour sample extraction OD=1.94, 12:55- J45120 17 hour sample extraction OD=2.12, 1:15- J45181 17 hour sample extraction OD=1.84
NOTE- I think that growing up the cultures in 125-mL flasks from the start altered the growth curves
3. Confirm flight arrangements for conference- DONE
4. Run samples above on GC
5. 3:00- Remove J45995, J45996, R0040.E0840, B0015 cultures and dilute them 1:250 at 220 RPM at 37C for 4 hours
6. GC extract cultures from last night adding internal (hexyl acetate before extraction) and external standards (octyl acetate after extraction) for J45200 and J45250 (just want to see if we see a peak for J45600 and J45900)- DONE
- Note: As Alex suggested I will dilute the J45200 and J45250 1/5 since we got a boatload last time
- We want 200 ppm internal and external standard.
- For hexyl acetate, .87 g/mL/.0001 g/mL= 1:8700 dilution; in 20 mL, 20 mL/8700 = 2.3 uL; add 2.3 uL to cultures before extraction
- For oxyl acetate, .856 g/mL/.0002 g/mL= 1:8560 dilution; in 1 mL, 1 mL/8560 = .1168 uL; Make a 1:10 dilution in heptane and add 1.168 uL to 1 mL sample
3. Make dilutions of isoamyl acetate in heptane (will add external and internal standards later if we see that they do not coelute with isoamyl acetate)- DOEN
- Calculations:
500 ppm----> .876 g/mL/.0005 g/mL= 1:1752 dilution; in 2 mL heptane, 1.142 uL isoamyl acetate
200 ppm----> 1.2 mL heptane + .8 mL 500 ppm isoamyl acetate
100 ppm----> 1 mL heptane + 1 mL 200 ppm isoamyl acetate
25 ppm----> 1.5 mL heptane + .5 mL 100 ppm isoamyl acetate
5 ppm----> .8 mL heptane + .2 mL 25 ppm isoamyl acetate
7. Prepare slides for lab meeting 5-7
8. 7:00- Dilute J45995, J45996, R0040.E0840, B0015 cultures and run them at 220 RPM at 37C for 50 mins
9. Extract 200 uL from each culture and run on the plate reader
10. Email Lily about GC- DONE
