Biology Reference
In-Depth Information
mutations on microtubule dynamics and interactions with specific regulatory pro-
teins, to distinct microtubule-dependent processes.
22.1
REAGENTS AND EQUIPMENT
General equipment
Temperature-controlled waterbath
0
C incubator and shaker
Thermocycler
Tabletop centrifuge
Tabletop microfuge
Spectrophotometer
Dissection microscope with micromanipulator
Replica plate apparatus with sterile velvet towels
Fluorescence microscope equipped with a 63
high N.A. objective, CCD
camera
Microscope slides and coverglass
SDS-PAGE/western blot apparatus and supplies
Multichannel pipette or “frogger”
Sterile wooden toothpicks
15-ml Conical tubes
Culture tubes and flasks
PCR tubes
96-Well plates
General reagents
Yeast-extract peptone dextrose (YPD) media (1% yeast extract, 2% peptone,
2% glucose)
YPD plates (YPD, 2% agar)
Synthetic complete (SC) media (0.67% yeast nitrogen base without amino
acids, 2% glucose, 0.2% appropriate amino acid mix (Sunshine Science
Products, CA))
SC plates (SC, 2% agar)
Sporulation plates (1% potassium acetate, 0.1% Bacto-yeast extract, 0.05%
glucose, 2% agar)
YNB plates (0.67% yeast nitrogen base without amino acids, 2% glucose,
2% agar)
For more details on preparing yeast media, see
Treco and Lundblad (2001)
Single-stranded salmon sperm carrier DNA; for preparation protocol, see
Becker and Lundblad (2001)
Adenine, NaOH, KCl, lithium acetate (LiAc), glycerol, polyethylene glycol
MW 3350, benomyl, DMSO (Sigma, St. Louis, MO)
Restriction enzymes (see
Table 22.1
)
TE buffer (10 mM Tris, 1 mM EDTA, pH 8)
