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Molecular Biology Test 1 > Notes 3 and 4 > Flashcards

Notes 3 and 4 Flashcards

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1
Q

Southern blotting steps

A
  1. DNA run onto gel: looks like smear
  2. Filter paper holds DNA
  3. Use probe thats highly specific to your gene of interest
  4. Wash away unbound probe
  5. Develop autoradiogram and expose to xray film
  6. Capture image on film
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2
Q

Type of filter paper used in Southern bloting

A

nitrocellulose

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3
Q

In Situ Hybridization (4 steps)

A
  1. Gently denature chromosomes that are in metaphase
  2. Use probe labeled with fluorescent dye
  3. Denature and hybridize
  4. Locate where it is on chromosome “the site”
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4
Q

Northern blotting

A

Transfer of RNA from a gel to a membrane

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5
Q

Western blotting steps

A
  1. Separate proteins on SDS gel
  2. Transfer to nitrocellulose membrane using electrotransfer
  3. This is called: Polyacrylamide gel electrophoresis
  4. Use antibodies to give specificity to proteins
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6
Q

What does SDS do?

A

coats proteins with a negative charge

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7
Q

How are proteins separated by polyacrylamide gel electrophoresis

A

Separated on size

No shape or charge component

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8
Q

Yeast model organisms

A

Saccharomyces cerevisiae

Schizosaccharomyces pombe

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9
Q

Budding yeast

A

Saccharomyces cerevisiae

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10
Q

Fission yeast

A

Schizosaccharomyces pombe

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11
Q

Slime Mold

A

Dictyostelium discoideum

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12
Q

Nematodes

A

Caenorhabditis elegans

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13
Q

Fruit fly

A

Drosophila melanogaster

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14
Q

Silk worm

A

Bombyx morii

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15
Q

Frogs

A

Rana pipens

Xenopus laevis

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16
Q

Typical dissection frog

A

Rana Pipens

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17
Q

African clawed frog

A

Xenopus laevis

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18
Q

Zebrafish

A

Danio rerio

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19
Q

Mouse

A

Mus musculus

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20
Q

Vascular plants

A

Arabidopsis thaliana

21
Q

What’s special about wood frogs?

A

They’re very freeze tolerant and will freeze solid in winter

22
Q

How do they prevent ice from damaging them?

A

Have polysaccharides to make sure ice doesn’t damage them

23
Q

when the sperm and egg fuse its called

A

syngamy

24
Q

When does the first cleavage occur?

A

90 minutes

25
Q

When does the gastrula occur?

A

10 hours

20,000 cells

26
Q

When does division slow (cell number)

A

4,096 cells

27
Q

Upper side of the egg is called

A

Animal hemisphere

28
Q

Lower side of the egg is called

A

Vegetal hemisphere

29
Q

When it divides to four cells, those cells are called…

A

blastomeres

30
Q

At 4,000 cells there’s the ______ stage

A

Blastula

31
Q

What forms inside the blastula

A

blastocoel

32
Q

What forms after the blastula?

A

Gastrula

33
Q

When does the notochord and neurula form?

A

about 13 hours

34
Q

When does a tail bud form?

A

1 day

35
Q

When does a tadpole form

A

3 days

36
Q

What is the mid blastula transition?

A

Where RNA synthesis occus and germ layers can develop

37
Q

Mesoderm

A

muscle, bone

38
Q

Ectoderm

A

skin, nervous system

39
Q

Endoderm

A

gastro lining

40
Q

How is the frog cell cycle abbreviated?

A

only has S & M stage

Replicate, divide

41
Q

Rapidly proliferating cells

A

Blastomeres

42
Q

5 questions of inquiry

A
  1. Observation and questions
  2. hypothesis
  3. testing the hypothesis
  4. Results
  5. Conclussion
43
Q

Steps of flow cytometry

A
  1. Prepare embryonic nuclei from blastula
  2. Stain with propidium iodide
  3. excite with UV
  4. Quantify fluorescence by flow cytometry to determine DNA content
44
Q

How is the mesoderm formed?

A

Contacting animal and vegetal layers

45
Q

How do we break down cell layer?

A

CMFM (calcium magnesium free medium)

46
Q

What are dispersed cells?

A

They aren’t touching

47
Q

What are dissociated cells?

A

They’re touching, in a pile

48
Q

What happened with dispersed cells vs. dissociated?

A

Dispersed cells lost their way and only stayed in S phase. Dissociated were similar to an intact embryo and still developed.

Molecular Biology Test 1 (4 decks)

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