Difference between revisions of "Will Notebook2"
Wideloache (talk | contribs) |
Wideloache (talk | contribs) |
||
| Line 9: | Line 9: | ||
--> | --> | ||
[[Will_DeLoache Notebook|Back to Main Page]] | [[Will_DeLoache Notebook|Back to Main Page]] | ||
| − | ==[[User:Wideloache|Wideloache]] | + | ==[[User:Wideloache|Wideloache]] 20:36, 13 January 2009 (EST)== |
| + | Today was my first day back in the lab of the semester. The first thing that I needed to do was to get the two pLux promoters that had been synthesized by GeneArt into pSB1A2. I had done an (E/P) digestion of pLux' and pLuxLas last semester that seemed to be successful. However, when I tried to transform into pSB1A2, I had a low colony yield. I decided to make a fresh stock of pSB1A2 digested with E/P. I used the part J61002-J23100. Cutting this with E/P woudl result in a pSB1A2 vector with a RFP expression insert. In this way I could screen out unsuccessful ligations. In addition to running this digestion O/N, I also redigested pLux' and pLuxLas with E/P from the minipreps of pLuxLacIX86 and pLuxLasLacII12X86. Tomorrow i will run these digestions next to my gel purifications from last semester to insure that those are the correct length insert. | ||
| + | |||
| + | <pre> | ||
| + | Make GenR plates (15 ug/mL) | ||
| + | Miniprep pLas' and pLasLux | ||
| + | Get GFP plasmid to put 4 promoters in front of. | ||
| + | </pre> | ||
| + | === === | ||
Revision as of 01:36, 14 January 2009
Wideloache 20:36, 13 January 2009 (EST)
Today was my first day back in the lab of the semester. The first thing that I needed to do was to get the two pLux promoters that had been synthesized by GeneArt into pSB1A2. I had done an (E/P) digestion of pLux' and pLuxLas last semester that seemed to be successful. However, when I tried to transform into pSB1A2, I had a low colony yield. I decided to make a fresh stock of pSB1A2 digested with E/P. I used the part J61002-J23100. Cutting this with E/P woudl result in a pSB1A2 vector with a RFP expression insert. In this way I could screen out unsuccessful ligations. In addition to running this digestion O/N, I also redigested pLux' and pLuxLas with E/P from the minipreps of pLuxLacIX86 and pLuxLasLacII12X86. Tomorrow i will run these digestions next to my gel purifications from last semester to insure that those are the correct length insert.
Make GenR plates (15 ug/mL) Miniprep pLas' and pLasLux Get GFP plasmid to put 4 promoters in front of.
