Will Notebook2
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.
