Lab 4 Flashcards
1
Q
What is the purpose of adding breaking buffer?
A
thaw the pellet
2
Q
Why do we vortex and incubate at 37 C?
A
improves the efficiency of lysing
3
Q
The supernatant yielded after breaking buffer/incubation/first round of centrifugation is called
A
the GFP crude extract (GCE)
4
Q
What is the bed volume?
A
volume of the matrix (beads) - 15 mL of a 50% slurry would yield 7.5 mL bed volume
5
Q
What is “pre-equilibrating the column?”
A
adding 10-fold breaking buffer to wash off the 20% ethanol
