IGEM:Harvard/2008/Lab Notebooks/DailyBook/Week13/Chemical and Light: Difference between revisions
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=09/30 | =Colony PCR 09/30= | ||
Should grow only on carb | Should grow only on carb | ||
*1-30: 20, 23, 25 (picked all three) --> P38+130 | *1-30: 20, 23, 25 (picked all three) --> P38+130 | ||
Line 15: | Line 15: | ||
=Amy's mutants= | =Amy's mutants= | ||
[[Image:2008-09-30_amymut.jpg]] | [[Image:2008-09-30_amymut.jpg]] | ||
=RE digests 10/01= | |||
Numbers refer to the master plate from 09/30 colony PCR. We digested with EcoRI, SpeI, and XmnI. | |||
*P38+130 #20 | |||
*P39+130 #31 | |||
*P39+130 #39 | |||
The expected band sizes are 2.2 kb (part), 1.6 kb and 0.4 kb (both vector backbone). | |||
=Ligations/transformations 10/03 = | =Ligations/transformations 10/03 = | ||
We ligated the following plasmids. The ligations were transformed into XL1-Blue. | We ligated the following plasmids. The ligations were transformed into XL1-Blue. | ||
* | *P108>131 | ||
* | *P38+51>131 | ||
* | *P122>131 | ||
* | *P38+130>101 | ||
* | *P130>133 | ||
*130> | *P130>134 | ||
There were two colonies on the P130>134 plate. These were grown up in liquid culture to be sent for sequencing. There were no colonies on the other plates. | |||
=Shewanella transformations 10/05= | |||
We ethanol precipitated the above ligations prior to electroporation of Shewanella. | |||
We transformed the following plasmids into wildtype Shewanella: | |||
*P130>134 | |||
*P38+51>131 | |||
*P130>133 | |||
*P122>131 | |||
*P38+130>101 | |||
*P108>131 | |||
*P138 #1 | |||
*P139 #1 | |||
*P139 #2 | |||
*P139 #3 | |||
*P139 #4 | |||
*P139 #5 | |||
*P140 | |||
*P142 | |||
We transformed the following plasmids into ΔmtrB Shewanella: | |||
We | *P130>134 | ||
*P38+51>131 | |||
*P130>133 | |||
*P122>131 | |||
*P38+130>101 | |||
*P108>131 | |||
*P138 #2 | |||
*P139 #1 | |||
*P140 | |||
*P142 |
Latest revision as of 15:55, 5 October 2008
Colony PCR 09/30
Should grow only on carb
- 1-30: 20, 23, 25 (picked all three) --> P38+130
- 31-60: 31-60 (picked 31, 35, 39, 42, 46, 50, 55, 60) --> P39+130
Should grow only on kan
- 61-90: none --> P116+130
- 91-120: none --> P117+130
- 121-150: none --> P108+130
--Upload picture of colony PCR gel--
Picked colony 20 (P38+130) and 31, 39, 60 (P39+130) and grew them up in liquid cultures (LB Amp).
Amy's mutants
RE digests 10/01
Numbers refer to the master plate from 09/30 colony PCR. We digested with EcoRI, SpeI, and XmnI.
- P38+130 #20
- P39+130 #31
- P39+130 #39
The expected band sizes are 2.2 kb (part), 1.6 kb and 0.4 kb (both vector backbone).
Ligations/transformations 10/03
We ligated the following plasmids. The ligations were transformed into XL1-Blue.
- P108>131
- P38+51>131
- P122>131
- P38+130>101
- P130>133
- P130>134
There were two colonies on the P130>134 plate. These were grown up in liquid culture to be sent for sequencing. There were no colonies on the other plates.
Shewanella transformations 10/05
We ethanol precipitated the above ligations prior to electroporation of Shewanella.
We transformed the following plasmids into wildtype Shewanella:
- P130>134
- P38+51>131
- P130>133
- P122>131
- P38+130>101
- P108>131
- P138 #1
- P139 #1
- P139 #2
- P139 #3
- P139 #4
- P139 #5
- P140
- P142
We transformed the following plasmids into ΔmtrB Shewanella:
- P130>134
- P38+51>131
- P130>133
- P122>131
- P38+130>101
- P108>131
- P138 #2
- P139 #1
- P140
- P142