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DNA Fragmentation and Cloning Vectors

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Year 2 Quiz on DNA Fragmentation and Cloning Vectors, created by gina_evans0312 on 29/12/2013.
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Quiz by gina_evans0312, updated more than 1 year ago
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Created by gina_evans0312 over 10 years ago
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Resource summary

Question 1

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

Question 2

Question
Non-random shearing involves the use of helicases
Answer
  • True
  • False

Question 3

Question
Which of the following cuts most frequently?
Answer
  • 4 cutters
  • 6 cutters
  • 8 cutters

Question 4

Question
A 4 cutter cuts a 4 bp sequence, and so on
Answer
  • True
  • False

Question 5

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

Question 6

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

Question 7

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

Question 8

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

Question 9

Question
Why is E.Coli most often used as a cloning vector?
Answer
  • 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

Question 10

Question
Bacteriophages can be used to transplant vectors into E.Coli
Answer
  • True
  • False

Question 11

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

Question 12

Question
Once inside the cell, the ssDONA of M13...
Answer
  • 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

Question 13

Question
TEVEN and Lambda enter the bacteria directly through the pore
Answer
  • True
  • False

Question 14

Question
What is the point of the resistance genes in this Generation 1 plasmid
Image:
Gen_1_Plasmid (image/jpg)
Answer
  • 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

Question 15

Question
What gene is disrupted in Gen 2 plasmids?
Image:
Gen_2_Plasmid (image/jpg)
Answer
  • The small subunit of LacZ
  • The large subunit of LacZ
  • Ampicillin
  • Tetracycline

Question 16

Question
Once the LacZ subunit has been assembled, what form does LacZ take?
Image:
Lac_Z (image/png)
Answer
  • Monomer
  • Dimer
  • Tetramer
  • Hexamer

Question 17

Question
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
Answer
  • True
  • False

Question 18

Question
Why is the dish impregnated with X-Gal?
Answer
  • 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

Question 19

Question
If the gene is successfully inserted, the colony will remain white
Answer
  • True
  • False

Question 20

Question
A Phagemid is virally inserted DNA in plasmid form
Answer
  • True
  • False

Question 21

Question
Why would a vector be given an M13 origin and inserted with M13?
Answer
  • 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

Question 22

Question
Cosmids are plasmids with...?
Answer
  • Cos lambda sites
  • Cos beta sites
  • Cos gamma sites

Question 23

Question
Which of the following has the largest limit?
Answer
  • BAC's
  • YAC's
  • MAC's

Question 24

Question
Why do you remove a phosphate from the ends of bacterial DNA using a phosphatase?
Image:
Remove_P (image/png)
Answer
  • To prevent it self ligating
  • To prevent it degrading
  • To prevent it getting destroyed

Question 25

Question
Why mix enzyme T4 and ATP together with the DNA to be inserted?
Answer
  • 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

Question 26

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