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Yeast PP 2

Submitted by mglater on Thu, 04/19/2018 - 19:26

Yeast were plated and each plate was mutagenized by Dr. Loomis by exposing the plate to UV radiation for 9 seconds via a UV light box. Cells that turned red were allowed to grow into larger colonies to be used for the experiment. Four different mutant strains were produced, two of mating type A and two of mating type alpha. Yeast labelled “A” were of the A mating type, while yeast labelled “B” were of the alpha mating type. A YED plate was set up in a grid to perform crosses (Figure 2). Each side consisted of one mating type of non-mutant yeast (HA0/HB0), a known Ade1 mutant (HA1/HB1), a known Ade2 mutant (HA2/HB2), and the two mutant strains for that mating type (MA1/2, MB1/2). After one day of growth, the yeast were crossed in a gridwise manner. Two days later, the yeast were replica plated to an MV plate and an MV+Adenine plate (Figure 3). After three additional days of incubation, the yeast colonies were observed. The full procedures followed for all steps can be found on Moodle.

 

Yeast

Submitted by mglater on Thu, 04/19/2018 - 19:25

Yeast were plated and each plate was mutagenized by Dr. Loomis by exposing the plate to UV radiation for 9 seconds via a UV light box. Cells that turned red were allowed to grow into larger colonies to be used for the experiment. Four different mutant strains were produced, two of mating type A and two of mating type alpha. Yeast labelled “A” are of the A mating type, while yeast labelled “B” are of the alpha mating type. A YED plate was set up in a grid to perform crosses (Figure 2). Each side consisted of one mating type of non-mutant yeast (HA0/HB0), a known Ade1 mutant (HA1/HB1), a known Ade2 mutant (HA2/HB2), and the two mutant strains for that mating type (MA1/2, MB1/2). After one day of growth, the yeast were crossed gridwise. Two days later, the yeast were replica plated to an MV plate and an MV+Adenine plate (Figure 3). After three additional days of incubation, the yeast colonies were observed. The full procedures followed for all steps can be found on Moodle.

 

Yeast

Submitted by mglater on Tue, 04/17/2018 - 15:18

The purpose of this study was to examine mutations in the yeast (Saccharomyces cerevisiae) adenine biosynthesis pathway. Adenine is one of the four nucleotides making up DNA, and thus is a vital compound for the survival of the yeast. The synthesis pathway of adenine has many steps, with different enzymes changing the compound until finally adenine is produced. The enzymes involved in these steps are named “Ade” followed by a number. This study examined mutation in the Ade1 and Ade2 enzymes. Ade2 converts the compound AIR into CAIR, and Ade1 converts CAIR into SAICAIR

Yeast

Submitted by mglater on Mon, 04/16/2018 - 20:11

To determine whether the mutation in a red yeast cell is in Ade1 or Ade2, the knowledge of complementation was used. The idea of complementation is that when two mutant haploid cells mate and produce a diploid, the ability of the diploid to produce functional, non-mutant proteins depends on whether the parent mutations were in the same gene or different genes. If the mutations were in the same gene, the diploid would inherit two dysfunctional alleles, and would therefore also be a mutant phenotype. However, if the mutations were in different genes, then the diploid would have one mutant allele and one functional allele for each of the mutant genes. The functional gene would be able to produce a functional product, and the diploid organism would not show the mutant phenotype. In this experiment, if the mutations of the haploid parents were both in Ade1 or Ade2, the diploid offspring would not have a functional copy of either enzyme, and thus still be red. If one mutation was in Ade1 and the other was in Ade2, the mutations would complement, and the diploid yeast would appear to be wild-type. Through complementation analysis, the unknown mutant gene in a red yeast colony can be determined.

 

Yeast PP

Submitted by mglater on Sat, 04/14/2018 - 13:11

Through complementation analysis, unknown mutations within the yeast (Saccharomyces cerevisiae) adenine biosynthesis pathway were identified. Studying the ability of unknown mutant colonies to produce successful colonies with known mutants revealed the identity of the unknown mutation as either a mutation in Ade1 or Ade2. Four mutant strains were examined, two strains of the A mating type and two strains of the alpha mating type. The A type yeast produced living colonies when crossed with a known Ade2 mutant as well as the unknown alpha type mutants. The alpha mutants produced living colonies when crossed with a known Ade1 mutant as well as the unknown A type. Using complementation analysis it was determined that both unknown A mutations were in the Ade1 gene and both unknown alpha mutations were in the Ade2 gene.

Yeast

Submitted by mglater on Wed, 04/11/2018 - 19:57

Through complementation analysis, unknown mutations within the yeast (Saccharomyces cerevisiae) adenine biosynthesis pathway were identified. Studying the ability of unknown mutant colonies to produce successful colonies with known mutants revealed the identity of the unknown mutation as either a mutation in Ade1 or Ade2. Four mutant strains were examined, two of the A mating type and two of the alpha mating type. The A type yeast produced living colonies when crossed with a known Ade2 mutant as well as the unknown alpha type mutants. The alpha mutants produced living colonies when crossed with a known Ade1 mutant as well as the unknown A type. Using complementation analysis it was determined that both unknown A mutations were in the Ade1 gene and both unknown alpha mutations were in the Ade2 gene.

 

Scavenger Hunt

Submitted by mglater on Sun, 04/08/2018 - 22:46

An extremely focused laser has a very strong electric field at its narrowest point, known as the “beam waist.” The particles are attracted to the strongest field, the center of the beam, which allows for the use of the laser to move particles around. Away from the center of the beam is higher potential, so the particles move to the lower potential in the center of the beam.

Proposal method PP

Submitted by mglater on Thu, 04/05/2018 - 19:06

The traps used will be a version of the pitfall trap described by Youngman et. al., 2009. The trap design is a cup firmly planted in a hole dug in the ground, with a second cup of the same rim width resting inside. In the interior cup, a small amount of ethanol will be placed to kill/preserve the specimens collected. Each team will set three traps at their chosen location, with each trap being at least twenty feet apart from the others.

 

Cell division

Submitted by mglater on Wed, 04/04/2018 - 21:23

A549, H460, H1650 are non-small cell lung cancer lines

  1. % mitotic cells compared to control through siRNA transfection

  2. Knockdown using lentiviral shRNA

  3. Western blot; cells synchronized at G1/S, released for varying times

    1. p-TBK1 activated 4 hours post release

    2. Maximal TBK1 phosphorylation= maximal Histone H3 phosphorylation

D) BX795 inhibits TBK1; control cells at 9 hours post release had pH3S10, but treated cells didn’t

E) TBK1 depleted by siRNA transfection, no pH3S10 (small band at 9hr, not mentioned in paper)

F) PLK1 is mitotic kinase, may be substrate of TBK1;Testing if overexpressing PLK1 can overpower depleted TBK1, but it can’t rescue from TBK1 depletion; TBK1 must target other proteins to modulate mitosis

 

Physics Lab

Submitted by mglater on Sun, 04/01/2018 - 22:47

We think the caliper measurement is better, as the double slit experiment introduces other variables and uncertainties which must be taken into consideration, such as accuracy of measuring th dark spots, and the ctual distance of the hair from the light and screen.

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