IGEM:MIT/2006/Notebook/2007-8-17

Things to Do Today

1. Make reservations for the plate reader (which is supposedly fixed)- DONE (5:00 PM tomorrow)

2. 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/.0002 g/mL= 1:4350 dilution; in 25 mL, 25 mL/4350 = 5.75 uL; add 5.75 uL to cultures before extraction

• For oxyl acetate, .856 g/mL/.0002 g/mL= 1:4280 dilution; in 1 mL, 1 mL/4280 = .2336 uL; Make a 1:10 dilution in heptane and add 2.336 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----> .6 mL heptane + .4 mL 500 ppm isoamyl acetate

100 ppm----> .5 mL heptane + .5 mL 200 ppm isoamyl acetate

25 ppm----> .75 mL heptane + .25 mL 100 ppm isoamyl acetate

5 ppm----> .8 mL heptane + .2 mL 25 ppm isoamyl acetate

4. Make LCs of J45995, J45996, B0015, and R0040.E0840- DONE (5:30 PM)

5. Flesh out next step for wintergreen time courses with advisors- DISCUSSED WITH JASON AND AUSTIN, SENT OUT EMAIL WITH PROPOSED PLAN (AWAITING RESPONSES)

6. Check sequencing results of J45250 1AT3- DONE (NO NEW MUTATIONS)

7. Make sure registration stuff for Microbial Genetics Conference gets done- WORKING ON IT, registered online for the time being

8. Add precursor to 25-mL J45250 1AT3 culture and see if it smells- DONE

9. Make a glycerol of J45250 1AT3- DONE

10. Streak out a master plate of J45250 1AT3- DONE