DNA Fragmentation and Cloning Vectors

Descripción

Year 2 Test sobre DNA Fragmentation and Cloning Vectors, creado por gina_evans0312 el 29/12/2013.
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Test por gina_evans0312, actualizado hace más de 1 año
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Creado por gina_evans0312 hace casi 11 años
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Resumen del Recurso

Pregunta 1

Pregunta
Name the two methods of physical fragmentation
Respuesta
  • Shearing- filtering through a fine needed
  • Sonication
  • Acidification
  • Boiling

Pregunta 2

Pregunta
Non-random shearing involves the use of helicases
Respuesta
  • True
  • False

Pregunta 3

Pregunta
Which of the following cuts most frequently?
Respuesta
  • 4 cutters
  • 6 cutters
  • 8 cutters

Pregunta 4

Pregunta
A 4 cutter cuts a 4 bp sequence, and so on
Respuesta
  • True
  • False

Pregunta 5

Pregunta
The staggered ends of two different enzymes can be ligated together, but afterwards cannot be cut by the same enzymes
Respuesta
  • True
  • False

Pregunta 6

Pregunta
When dealing with DNA-cutting enzymes, what does the word degenerate mean?
Respuesta
  • The ends they produce don't last long
  • They can cut more than one sequence
  • Where in the sequence they cut changes

Pregunta 7

Pregunta
What conditions are necessary for a partial digest?
Respuesta
  • Not enough time
  • Less enzyme
  • Wrong enzyme conditions
  • A four cutter

Pregunta 8

Pregunta
Why could a partial digest be done?
Respuesta
  • To get longer fragments and more likely preserve the gene
  • To save time
  • To prevent the DNA from degrading

Pregunta 9

Pregunta
Why is E.Coli most often used as a cloning vector?
Respuesta
  • It has a rapid generation time
  • Its DNA is easy to manipulate
  • Its DNA is easy to get hold of
  • They don't mutate much

Pregunta 10

Pregunta
Bacteriophages can be used to transplant vectors into E.Coli
Respuesta
  • True
  • False

Pregunta 11

Pregunta
How does M13 infect the male E.Coli?
Respuesta
  • Binds to pilus surface
  • Enters through bacterial pores
  • Direct injection of DNA

Pregunta 12

Pregunta
Once inside the cell, the ssDONA of M13...
Respuesta
  • Is convered to dsDNA that acts like a plasmid
  • Is integrated into the bacterial chromosome
  • Is cut up by restriction enzymes ans integrated into the bacterial chromosome

Pregunta 13

Pregunta
TEVEN and Lambda enter the bacteria directly through the pore
Respuesta
  • True
  • False

Pregunta 14

Pregunta
What is the point of the resistance genes in this Generation 1 plasmid
Respuesta
  • Tetracycline is interrupted- E.Coli with no resistance have had the recombinant plasmid inserted
  • The recombinant DNA is inserted at the EcrR1 site
  • E.Coli with ampicillin resistance have taken up a version of the plasmid

Pregunta 15

Pregunta
What gene is disrupted in Gen 2 plasmids?
Respuesta
  • The small subunit of LacZ
  • The large subunit of LacZ
  • Ampicillin
  • Tetracycline

Pregunta 16

Pregunta
Once the LacZ subunit has been assembled, what form does LacZ take?
Respuesta
  • Monomer
  • Dimer
  • Tetramer
  • Hexamer

Pregunta 17

Pregunta
For Gen 2 plasmid testing to work, the LacZ gene must be removed from the E.Coli, and a plasmid for the large LacZ subunit added
Respuesta
  • True
  • False

Pregunta 18

Pregunta
Why is the dish impregnated with X-Gal?
Respuesta
  • It goes from blue to white in the presence of LacZ
  • It goes from white to blue in the presence of LacZ
  • It goes from white to blue in the presence of the small subunit of LacZ

Pregunta 19

Pregunta
If the gene is successfully inserted, the colony will remain white
Respuesta
  • True
  • False

Pregunta 20

Pregunta
A Phagemid is virally inserted DNA in plasmid form
Respuesta
  • True
  • False

Pregunta 21

Pregunta
Why would a vector be given an M13 origin and inserted with M13?
Respuesta
  • The M13 phage will treat the plasmid as its own DNA and replicate it for secretion
  • The phage will insert the vector into the chromosome
  • The phage will prevent the bacteria from destroying the plasmid

Pregunta 22

Pregunta
Cosmids are plasmids with...?
Respuesta
  • Cos lambda sites
  • Cos beta sites
  • Cos gamma sites

Pregunta 23

Pregunta
Which of the following has the largest limit?
Respuesta
  • BAC's
  • YAC's
  • MAC's

Pregunta 24

Pregunta
Why do you remove a phosphate from the ends of bacterial DNA using a phosphatase?
Respuesta
  • To prevent it self ligating
  • To prevent it degrading
  • To prevent it getting destroyed

Pregunta 25

Pregunta
Why mix enzyme T4 and ATP together with the DNA to be inserted?
Respuesta
  • Adding AMP to ligated ends gives easily ligable high energy form
  • ATP & T4 remove the phosphates from the ends of the DNA to prevent self annealing
  • T4 can only ligate DNA in the presence of ATP

Pregunta 26

Pregunta
A vector must be used because E.Coli cannot take up linear DNA
Respuesta
  • True
  • False
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