IGEM:MIT/2006/Notebook/2006-8-11

To do

1. smell overnight IK smell testers and after making glycerols from regular batch then add precursor to the daytime smell testers
2. analyze sequencing from weds [full assemblies]
3. List Biobrick Molecules
   • Lets update our index with our numbering system and all the biobrick constructs we believe that we have (and then we can check them off as we get the sequence results back. Also, BO could you put in links to each part's registry page on the index!)
4. digest WGD with EP and run on a gel with 8/10 pUCP22:EP digest
5. ligate WGD into EP cut pUCP22 shuttle vector and transform into Top10 in the pm
6. autoclave waste
7. digest pCHBA with E and S and backbone with E and S and ligate
8. mutagenesis of BAT2, transformation of BAT2, ATF1, and THI3 mutagenesis
9. LCs of R0040.E0840 and osmY.E0840s
10. GC of smell tests
11. miniprep RBS.coding.terminators to inverters

Preliminary Smell Test: Indole KO Strain (AG)

Experiment description

• We would like to know the amount of time it takes to produce the final volatile compound. I added SA (2mM) and IA-OH (10mM) to 5mL pre-grown Indole KO cultures.

	J45100	J45120	J45170	J45200	J45220
15 min	smells pretty bad, i.e. like regular Indole KO cells	smells pretty bad, i.e. like regular Indole KO cells	smells pretty bad, i.e. like regular Indole KO cells	faint smell of banana; smell of precursor lingering	isoamyl alcohol greatly overpowering
30 min	slight hint of mint; normal IKO-smell prevalent	very slight hint of mint; normal IKO-smell prevalent	smells pretty bad, i.e. like regular Indole KO cells	noticeable smell of banana, stronger than at 15 min	isoamyl alcohol greatly overpowering
60 min
90 min
120 min
150 min