IGEM:MIT/2006/Notebook/2007-1-12

Check out the checklist for each part on Stephen's contributions page to see progress.

J45400 did not grow up on any plate again.

To Do List

1. Run a gel of the colony PCR products of J45800- DONE- all bad, redo colony PCR with different polymerase- DONE and make LCs of those colonies- DONE

2. Take out the LCs of the good colony PCR colonies from J45120- DONE, perform antibiotic test on them- DONE

3. Check sequencing results of J45250 and J45400- DONE

4. Make plates- DONE and redo antibiotic test of pBANANAs from Jason- DONE, J45120- DONE, J45400- DONE, and controls- DONE

5. Do a smell test of J45200 and J45250

6. Streak the plate of J45180 from multiple colonies onto a new plate to find isolated colonies, which can be used for colony PCR tomorrow- DONE

7. Find Indole-Knockout Cells and ask Kate about them- DONE

8. Grow LCs of Mach Is and pBANANAs- DONE

Immediate Plan

J45120

Miniprep, sequence, glycerol, and antibiotic test 3 LCs of the correct colony PCRs from J45120. Note that the original cells failed the antibiotic test.

J45180

Restreak the J45180 "many colonies" plate, colony PCR, and look for the right size bands.

J45200

Conduct a smell test

J45250

Analyze sequencing results -- '''DONE, looks good! differences: 78 t->c, 79 g->c, tgc insert between 81 and 82, 178 a -> c

J45320

Await completion of J45180/J45120

J45400

Analyze sequencing results -- '''DONE, looks good! not perfect, though, will specify mutations tomorrow'''

J45600 (pBANANA)

Repeat streak tests on single antibiotic plates, grow LCs from prior antibiotic tests, then miniprep, sequence, and glycerol

J45700

Just J45320

J45800

Redo colony PCR