IGEM:PennState/Labbook/NoahJohnson/2007-8-9

August 9, 2007


 * Ran PCR cleanup on the 8 PCR rxns that worked from yesterday (all except 0.1uL for Buffer#1). Combined all into one sample.
 * Restriction digest with EcoR1 and SpeI
 * Assumed we maxed out the spin column so 10ug DNA was eluded in 28uL EB Buffer leading to a DNA concentration of ~357ng/uL

Restriction Digest Protocol 1. Added to tubes: 2. Gingerly flick tube to mix 3. Spin down tubes briefly 4. Incubate at 37C in water bath for 1-4 hrs (1-2 hrs optimal)
 * 2uL DNA (~350 ng/uL)
 * 1uL 10x Buffer
 * 1uL BSA
 * 0.5uL each restriction enzyme
 * 5uL dH2O


 * Also doing restriction on P1010 w/ psB1A2 which will be the vector for the following ligation.
 * Started incubating at 12:45pm


 * Ran P1010 on a gel to isolate it from psB1A2
 * Seems the restriction didnt work as planned. Going to grow up P1010 in psB1A2 overnight and miniprep and restrict again in the morning.

9:30-5