IGEM:MIT/2006/Notebook/2007-1-12: Difference between revisions
No edit summary |
No edit summary |
||
(13 intermediate revisions by 2 users not shown) | |||
Line 1: | Line 1: | ||
''Check out the checklist for each part on Stephen's contributions page to see progress.'' | ''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''' | '''To Do List''' | ||
1. Run a gel of the colony PCR products of J45800- DONE | 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 |
Latest revision as of 19:06, 12 January 2007
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