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 -done
2. analyze sequencing from weds [full assemblies] veena and kate, afternoon
3. List Biobrick Molecules
4. add pchBA to registry
   • 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!)
5. digest WGD with EP and run on a gel with 8/10 pUCP22:EP digest digest started 12:30
6. ligate WGD into EP cut pUCP22 shuttle vector and transform into Top10 in the pm
7. autoclave waste
8. digest pCHBA with E and S and backbone with E and S and ligate digest started 12:30
9. mutagenesis of BAT2, transformation of BAT2, ATF1, and THI3 mutagenesis Bo and Stephen pm
10. LCs of R0040.E0840 and osmY.E0840s
11. GC of smell tests decided to run it when we have more controlled samples to run
12. miniprep RBS.coding.terminators to inverters digests of these started at 12:30

Preliminary Smell Test: Indole KO Strain (André)

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.
  • Cultures containing J45100 and J45220 were further incubated at room temperature (25C)
  • Cultures containing J45120, J45170, and J45200 were further incubated (with vigorous shaking) at 37C

	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 IA-OH lingering	IA-OH 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	IA-OH greatly overpowering
60 min	slight hint of mint; normal IKO-smell prevalent	noticeable mint smell; normal IKO-smell prevalent	smells pretty bad, i.e. like regular Indole KO cells	powerful banana scent	IA-OH greatly overpowering
90 min	slight hint of mint; normal IKO-smell prevalent	noticeable mint smell; normal IKO-smell prevalent	smells pretty bad, i.e. like regular Indole KO cells	strong banana scent	IA-OH greatly overpowering
120 min	slight hint of mint; normal IKO-smell prevalent	noticeable mint smell; normal IKO-smell prevalent	smells pretty bad, i.e. like regular Indole KO cells	strong banana scent	banana smell is faint; IA-OH smell prevalent
180 min