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Creating an E.Coli Library

Descripción

Year 2 Test sobre Creating an E.Coli Library, creado por gina_evans0312 el 29/12/2013.
gina_evans0312
Test por gina_evans0312, actualizado hace más de 1 año
gina_evans0312
Creado por gina_evans0312 hace casi 11 años
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Resumen del Recurso

Pregunta 1

Pregunta
Electroporations- passing electricity through a mix of DNA and bacteria causes DNA to be taken up
Respuesta
  • True
  • False

Pregunta 2

Pregunta
What does N stand for?
Image:
Library_Size (image/png)
Respuesta
  • The no of clones needed
  • P of getting a clone with the desired gene
  • Genome size

Pregunta 3

Pregunta
What are the meanings of i and g?
Image:
Library_Size (image/png)
Respuesta
  • i = Library size g= probability of getting desired gene
  • i = avg insert size g = genome size
  • i = probability of getting desired gene g = genome size

Pregunta 4

Pregunta
The larger the genome, the larger the library required to replicate it
Respuesta
  • True
  • False

Pregunta 5

Pregunta
When is cDNA used?
Respuesta
  • When an organism has a lot of non-coding DNA
  • When an organism is very small
  • When the DNA is degraded

Pregunta 6

Pregunta
How is cDNA made?
Respuesta
  • Converted from mRNA
  • Converted from snoRNA
  • Converted from siRNA

Pregunta 7

Pregunta
Less than 1% of the genome is mRNA, but it's exclusively expressible genes
Respuesta
  • True
  • False

Pregunta 8

Pregunta
What is required to make the first RNA-DNA hybrid?
Image:
Hybrid (image/png)
Respuesta
  • dNTP's
  • Primer
  • Reverse Transcriptase

Pregunta 9

Pregunta
What are the two methods of degrading the RNA part?
Respuesta
  • Alkylisation (RNA is less stable)
  • RNA digestion through RNAase H
  • Boiling
  • Restriction enzymes

Pregunta 10

Pregunta
Which method was used to get the following DNA strand?
Image:
Alkali_Degrading (image/png)
Respuesta
  • Alkali degrading
  • Boiling
  • Shaking
  • RNAase H

Pregunta 11

Pregunta
What is required to get from the former to the latter?
Image:
Alkali_Degrading (image/png)
Respuesta
  • dNTP's
  • DNA Polymerase
  • Primer
  • Reverse transcriptase

Pregunta 12

Pregunta
What method was used to degrade the RNA into the following?
Image:
RNAase_H (image/png)
Respuesta
  • RNAase H
  • Alkylation
  • Boiling
  • Sonication

Pregunta 13

Pregunta
What is needed to get from the former to the latter?
Image:
Final_Product (image/png)
Respuesta
  • dNTP's
  • DNA Polymerase H
  • RNAase
  • rNTP's

Pregunta 14

Pregunta
When creating cDNA, a little information is always lost from the 3' end
Respuesta
  • True
  • False

Pregunta 15

Pregunta
How do we prevent this loss of information?
Respuesta
  • After degrading the RNA, add a homopolymer (polyC) tail to the 3 end
  • Using Terminal Transferase
  • Before degrading the DNA, degrade the Poly A tail

Pregunta 16

Pregunta
Then in second strand synthesis add ...
Respuesta
  • dNTP's
  • DNA polymerase
  • Oligo-dG primer

Pregunta 17

Pregunta
Enriching desired DNA means creating/finding more of it
Respuesta
  • True
  • False

Pregunta 18

Pregunta
Selective Hybridisation removes cDNA's common to more than one tissue
Respuesta
  • True
  • False

Pregunta 19

Pregunta
Tissue Specificity is useful if the gene is only expressed in a certain tissue
Respuesta
  • True
  • False

Pregunta 20

Pregunta
What is Normalisation?
Respuesta
  • Melt, anneal and isolate ssDNA
  • Use restriction enzymes to repeatdely cut and ligate DNA
  • Use DNA helicase to continually replicate DNA

Pregunta 21

Pregunta
How does Normalisation work?
Respuesta
  • Repetitive DNA binds to other repetitive DNA
  • Leaving only unique DNA single stranded
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