Question | Answer |
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) |