Cell and Molecular Biology Lab Final Exam Review Notes

In our transformation experiment, what were the two marker genes? Which one was the 
selectable marker? Which is the reporter marker? 

Greem florescent proteins (gfp is the reporter marker) and bata lactonase (selectable marker)

2. What is a plasmid, and why did we use it in our transformation?

Extrachromosomal circular DNA (green pea circular plasmid DNA) that is able to replicate independently of the bacterial genome. Used to amplify and transfer certain genes into organisms as a vector

What was the phenotype of successful transformants? What were the four treatments 
(plates) we used as a class? Which plate would you look at first to determine whether your 
transformation was successful?

Colonial growth showing ampicillin resistance and glows green. 4 Treatments: 
- LB-plasmid/LB+plasmid 
- LB/amp-plasmid 
- LB/amp+ plasmid(look here first as should show wanted phenotype)

Transformation is rare – what is the rate of successful transformation?

1 in 1,000

Why is it more difficult to successfully transform eukaryotes?

Because in akaryotes not only do you have to get the DNA across the plasmid membrane but also into the nucleus whereas bacteria will just express plasmid DNA (eukaryotes won't)

What step in the transformation procedure is the most critical to time correctly?

The 90s heat shock

Why do we incubate the colonies and plasmid in CaCl2?

That incubation is intended to give the plasmid DNA as close as possible to the plasma membrane or the cell wall of the bacteria

What genes did we amplify in the GMO lab and why?

CamV promoter and NOS terminator  

From what organisms are the GM genes derived?

CamV promoter derived from Cauliflower mosaic virus and NOS terminator derived from Agrobacterium Tumefaciens 

When does crossing over occur?

Prophase 1

When does independent assortment occur?

Metaphase one of meiosis

When does a synapsis form?

Prophase 1 of meiosis

When do homologous chromosomes separate?

Anaphase 1

When do tetrads line up at the middle of the spindle?

Metaphase 1

What is the chromosomal makeup of each daughter cell after meiosis I? After meiosis II?

Haploid, haploid

What happens during the different stages of the cell cycle?

Be able to recognize cells in different stages of mitosis or meiosis based on the number and arrangement of chromosomes.