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 standard later if we see that it does not coelute with isoamyl acetate)

4. Make LCs of J45995, J45996, B0015, and R0040.E0840 at the end of the day

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