IGEM:Harvard/2008/Lab Notebooks/DailyBook/Week13/Chemical and Light

=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