IGEM:MIT/2006/Notebook/2007-1-13

Results from Yesterday/Future Plans

J45120- All three clones passed the antibiotic test, meaning that we have isolated colonies with just the one plasmid in it. We will smell test tonight and then miniprep tomorrow morning if they work. Set up smell tests of M1, M2, and M4 (all J45120); made 10 mL cultures w/ 40uL SA + controls, to be smelled tomorrow -VV

J45180- I am colony PCRing those today. I also am making LCs of them today. They will be run on the gel along with the colony PCRs of J45800 from yesterday. expected length on gel: 2520 bases

J45200- cells aren't grown up enough to effectively see if they smell like banana. just checked them; they do smell banana-ish (but a little isoamyl alcoholish too, i thought; they're still growing -VV

J45250- cells smell like banana. YEH!!!! yes, they def smell : ) -VV

J45400- still will not grow up on AMP/KAN, AMP/TET, or AMP/CHL but sequencing results are correct.

pBANANA- pBANANA 10 and 12 grows on C and K and not A and T, Mach I grows on T and K and not A and C, controls work. This is the same result as before, and I used different plates. So, it seems that my results have to be correct. I will miniprep these tomorrow to submit for sequencing Monday.

J45800- These will be run on the gel with the J45180 colony PCRs later in the day. expected length on gel: 4480 bases

GELS RUN TODAY (sorry the images are huge)

Gel 1: lanes (left to right) = 2-log ladder, 181, 182, 183, 184, 185, 186, 187, 188, 189

Gel 2: lanes (left to right) = 2-log ladder, 1810, 1811, 1812, 121, 122, 123, 124, 125, 126

Gel 3: lanes (left to right) = 2-log ladder, 127, 128, 129, 241, 242, 243, 244, 245, 246

Gel 4: lanes (left to right) = 2-log ladder, 247, 248, 249