Wisconsin-Madison/17 June 2009

June 17, 2009
Chris and I were in lab today working on the digestion/ligation. The ligation should be finished tomorrow morning, and whoever arrives first should transform the product (located in the PCR machine at 16 degrees C) into competent DH10B cells. The spec was giving us inconsistent readings, so we are uncertain the DNA concentrations were as needed for ligation. GFP, gssdmt, and the triple transformation were inoculated and need to be minipreped.

TW, if we want to do double digestion, the New England Double Digestion Finder suggested we used NE Buffer EcoRI +BSA for EcoRI and SpeI, and NEBuffer 3 +BSA for XbaI and PstI, both at 3

After ligation, do we need to transform the new BioBrick plasmids into DH10B and miniprep them again?