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BZ 310- Lab Quiz 1

4 types of microscopy used in lab bright field, phase contrast, dark field, kohler
Kohler provides best possible resolution (can be used w/ all 3)
dark field is useful for motile cells like sperm
the india ink showed us brownian motion
what did we view under phase contrast? pond water
what did we view under bright field? onion skin
white pipette tips are for P1000
yellow pipette tips are for P20 and P200
P10 "0650" 6.5 uL
P20 "178" 17.8 uL
P200 "150" 150 uL
P1000 "0670" 0.67 mL or 670 uL
what were the 2 expected locations of the GFP? nucleus and cytoplasm/bud
saccharomyces cerevisiae means "sugar splitter of beer"
yeast is the generic term for a single-celled fungus
haploid rounded, divide by budding, die under nutritional stress
diploid (shape etc) larger/oval, divide by budding, nutritional stress= meiosis to form 4 spores
two unknown strains YPH499 (a), YPH500 (alpha)
alpha strain Y43
a strain Y44
diploid DY (cross between YPH 499 and Y43)
YPD rich medium (yeast extract, peptone, dextrose)
SD minimal medium (synthetic w/ dextrose)
6 sections of the plate unknown, Y44+unknown, Y43+unknown, Y43, Y44, DY
alpha express... alpha transcription factor from MAT locus
a express 'a' transcription factor from MAT locus
3 DNA primers MAT forward primer, a reverse primer, alpha reverse primer
what was in the master mix? buffers (w/ dNTPs), primers, taq polymerase
BLAST basic local alignment search tool
PCR 1x of what? beginning: 3 min 96C (denaturing) end: 3 min 72C (extension)
PCR 35x of what? 15 sec 96C (denaturing) 30 sec 55C (annealing) 2 min 72C (extension)
DNA is ___ charged and will move to the ____ electrode negatively, positive
two types of gels in electrophoresis agarose and polyacrylamide
mobility in agarose is determined by size AND shape (not mass), but linear PCR is basically based on weight
agarose gel solution 1% agarose, EtBr, TBE buffer
DNA loading dye 10% glycerol and bromophenol blue
electrophoresis, how long and how many volts? 100 V for 30 minutes
two ways yeast cells divide budding (2 diploid), meiosis (4 haploid)
specialized cell junctions cardiac muscle create wave of depolarization
no PCR product, why? wrong strain, ran off the gel
positive control yeast diploid strain
why are shmoos formed? going toward higher pheromone concentration, will form a diploid
why did we do complementation with yeast? to determine mating type (needed to complement missing genes on YPD plate for growth to occur)
Created by: melaniebeale