http://2008.igem.org/wiki/index.php?title=Team:Harvard/Dailybook/Week13/Chemical_and_Light&feed=atom&action=historyTeam:Harvard/Dailybook/Week13/Chemical and Light - Revision history2024-03-29T11:07:43ZRevision history for this page on the wikiMediaWiki 1.16.5http://2008.igem.org/wiki/index.php?title=Team:Harvard/Dailybook/Week13/Chemical_and_Light&diff=69202&oldid=prevJoyy: New page: =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-...2008-10-26T19:22:41Z<p>New page: =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-...</p>
<p><b>New page</b></p><div>=Colony PCR 09/30=<br />
Should grow only on carb<br />
*1-30: 20, 23, 25 (picked all three) --> P38+130<br />
*31-60: 31-60 (picked 31, 35, 39, 42, 46, 50, 55, 60) --> P39+130<br />
<br />
Should grow only on kan<br />
*61-90: none --> P116+130<br />
*91-120: none --> P117+130<br />
*121-150: none --> P108+130<br />
<br />
--Upload picture of colony PCR gel--<br />
<br />
Picked colony 20 (P38+130) and 31, 39, 60 (P39+130) and grew them up in liquid cultures (LB Amp).<br />
<br />
=Amy's mutants=<br />
[[Image:2008-09-30_amymut.jpg]]<br />
<br />
=RE digests 10/01=<br />
Numbers refer to the master plate from 09/30 colony PCR. We digested with EcoRI, SpeI, and XmnI.<br />
*P38+130 #20<br />
*P39+130 #31<br />
*P39+130 #39<br />
<br />
The expected band sizes are 2.2 kb (part), 1.6 kb and 0.4 kb (both vector backbone).<br />
<br />
=Ligations/transformations 10/03 =<br />
We ligated the following plasmids. The ligations were transformed into XL1-Blue.<br />
<br />
*P108>131<br />
*P38+51>131<br />
*P122>131<br />
*P38+130>101<br />
*P130>133<br />
*P130>134<br />
<br />
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.<br />
<br />
=Shewanella transformations 10/05=<br />
We ethanol precipitated the above ligations prior to electroporation of Shewanella.<br />
<br />
We transformed the following plasmids into wildtype Shewanella:<br />
*P130>134<br />
*P38+51>131<br />
*P130>133<br />
*P122>131<br />
*P38+130>101<br />
*P108>131<br />
*P138 #1<br />
*P139 #1<br />
*P139 #2<br />
*P139 #3<br />
*P139 #4<br />
*P139 #5<br />
*P140<br />
*P142<br />
<br />
We transformed the following plasmids into ΔmtrB Shewanella:<br />
*P130>134<br />
*P38+51>131<br />
*P130>133<br />
*P122>131<br />
*P38+130>101<br />
*P108>131<br />
*P138 #2<br />
*P139 #1<br />
*P140<br />
*P142</div>Joyy