IGEM:The Citadel/Notebook/Notes Patrick

Friday, June 25, 2010

11:14am

Removed inoculated conical tubes from yesterday from incubator.

Removed the following plates from yesterday from incubator and placed into 4°C.
Agar in some plates was deformed and not completely dry
KEPT THE FOLLOWING PLATES (Listed according to quality of gel, not necessarily colony formation):
BBa_F1610 [100%]
- Completely dry.
- Many cells, one large cluster.
BBa_J45120 [100%]
- Completely dry.
- Many cells. Some formation along rim of agar.
BBa_R0063 [25%]
- Completely dry.
- Average colonies.
BBa_R0063 [25%]
- Completely dry.
- Average colonies.
BBa_R0040 [25%]
- Almost completely dry.
- Very few colonies.
BBa_F2620 [25%]
- Completely dry.
- Few colonies, but good ones.
BBa_F1610 [50%]
- Completely dry.
- Normal colonies.
BBa_E0240 [50%]
- Completely dry.
- Very few colonies. Colonies along rim.
BBa_I0462 [100%]
- Some liquid. Some agar on cover.
- Many colonies.
BBa_I0462 [50%]
- Some liquid.
- Normal/average colonies.
BBa_I0462 [25%]
- Slightly more liquid. Liquid agar on 1/2 of rim circumferance.
- Few colonies. Colonies along rim.
BBa_F2620 [50%]
- Liquid agar on 1/3 of rim.
- Average colonies.
BBa_J45120 [25%]
- Few colonies. Small colonies. Bacterial smear covering 1/12 of plate.
BBa_C0160 [25%]
- Liquid agar on 1/2 of rim.
- Very few colonies, but good ones.
BBa_B0015 [25%]
- Liquid agar on entire rim.
- Average colony formation.
DISCARDED THE FOLLOWING PLATES
BBa_J45200 [25%]
- Too much liquid agar on cover. Unsolidified gel.
- No visible colonies.
BBa_C0160 [50%]
- Too much liquid agar on cover. Unsolidified gel.
- Irregular colony formation across unsolidified gel.
BBa_B0015 [50%]
- Completely unsolidified gel.
- Bacteria formation on cover. Irregular colonies.
BBa_J45120 [50%]
- Too much liquid agar on cover. Unstable gel.
- Carpet style formation of bacteria.
BBa_J23102 [50%]
- Too much liquid agar on cover. Unstable gel.
- Bacteria formation on cover.
BBa_J45200 [50%]
- Too much liquid agar on cover. Unsolidified gel.
- No visible colonies.
BBa_R0062 [50%]
- Too much liquid agar on cover. Unstable gel.
- Carpet style formation of bacteria.
BBa_R0062 [50%]
- Agar dripping onto cover, taking colony formations with it.
BBa_E0240 [25%]
- Completely dry. Agar slightly unstable.
- Would be difficult to isolate from.
NOTES
Need to replate BBa_J45200 [25%], [50%].
Need to replate BBa_J23102 [25%], [50%].
Need to replate BBa_B0015 [50%].
Need to replate BBa_C0160 [50%].
Need to replate BBa_R0062 [25%], [50%].
Need to replate BBa_J4512 [50%].
Need to replate BBa_E0240 [25%].
In the future:
- Do not reheat agar solution in water bath during plate pouring.
- Ensure that plates have adequate time to dry completely.

12:45pm

Prepare LB-Miller (Amp) broth for x24 5mL liquid cultures
Prepare LB-Miller (Amp) agar 500mL
- Therefore, use total 1.24mL Amp (50μL/mL stock)

1:55pm

Plated BBa_I20260 [100%], [50%], [25%] Placed plates into 37°C incubator.

Tuesday, June 29, 2010

~12:30pm

Streak plated from isolations of the following parts and placed into incubator:
BBa_R0063
BBa_I20260

~3:00pm

Poured 33 LB-Miller (Kan) plates and 13 LB-Miller (Amp) plates

4:33pm

Inoculated the following and placed into incubator for miniprep tomorrow morning:
BBa_I20260 in 5mL LB Miller (Kan) broth
BBa_J45120 in 5mL LB Miller (Amp) broth
BBa_I0462 in 5mL LB Miller (Amp) broth

Wednesday, June 30, 2010

9:45am

Yesterday's plating of BBa_I20260 failed.
Replated BBa_I20260
Placed into incubator and 9:59am.

Wednesday, July 08, 2010

9:30am

Worked on theory for most of morning and afternoon.

2:00pm

Inoculated 80μL of LB-Miller (Amp) broth with following parts and placed into incubator.
I13521
J23101
J45120
I0462
I20260
(Cultures will be removed tomorrow morning and added to 40μL of glycerol(50%) and stored at -80°C.)

Thursday, July 15, 2010

9:30am

Brian made SOC.
Humter prepared LB-Miller (Amp) plates.

5:15pm

Inoculated 10mL of LB-Miller (No antibiotic) broth with parts from 07-14 MIT shipment and placed into incubator.
Inoculated 3mL of LB-Miller (no antibiotic) broth with Top 10 E.coli and placed into incubater. (For use in growth measurements tomorrow.)

Friday, July 16, 2010

9:00am

Hunter performed miniprep on cultures that were inoculated with 07-14 MIT shipment parts yesterday.
Brian electroporated the following parts:
- I746350
- I746351
- I746361
- I746362
- I746360
- I746364
- I746365 Note: "2.49 capacitance this time. Normally 2.50."
- I746352
- B0034 (Electroporated with our self-made SOC, as were all parts above.)
- B0034 (Electroporated with stock SOC.)