Introduction To Molecular Biology an der Universität Für Bodenkultur Wien | Karteikarten & Zusammenfassungen

Lernmaterialien für Introduction to Molecular Biology an der Universität für Bodenkultur Wien

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TESTE DEIN WISSEN

Watson and Crick?

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TESTE DEIN WISSEN
  • the composition of DNA varies from one species to another
  • total amounts of pyrimidine equal to the amounts of purine: A+G=C+T
  • after Frnklin and Gosline made a xray diffraction analysis they comined the information
    • Nucleotides from a linear chein, linked by phospodiester bonds
    • DNA froms a holical double strand
    • antiparallel organization
    • minor and major groove
    • 1 double helix turn - 10 bp
Lösung ausblenden
TESTE DEIN WISSEN

Identification of recombinant clones (electroporation)?

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TESTE DEIN WISSEN
  • after electroporation
    • many Ecoli will have taken up one plasmid
    • very few will contain 2 oder more
    • many Ecoli cells without plasmid DNA are present
  • each plasmid molecule carries the selection marker gene (pET11a -> ampicillin)
  • on a medium containing ampicillin only Ecoli that carry a plasmid are able to grow
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TESTE DEIN WISSEN

Human genome size?

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TESTE DEIN WISSEN

3234 Mbp

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Histone Code?

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TESTE DEIN WISSEN

numerous possible combinations

  • various effects on gene expression
  • highly dynamic and reversible
  • interaction between chromatin remodelling, modification histone variants -> impacts on gene expression


Lösung ausblenden
TESTE DEIN WISSEN

PCR History

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TESTE DEIN WISSEN
  • first proposed by Khorana in 1970
  • made public by Kary Mullis in 1985
  • method of the year in 1989
  • nobel prize in 1993
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TESTE DEIN WISSEN

DNA sequencing?

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TESTE DEIN WISSEN
  • 2 types of DNA sequencing
    • Dideoxy cycle DNA sequencing:
      • for sequencing single clones
    • High troughput, next generation DNA sequencing:
      • for genome and transcriptome sequencing
Lösung ausblenden
TESTE DEIN WISSEN

qPCR?

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TESTE DEIN WISSEN
  • Aim: detection and quantitation of specific DNA in samples
  • medical tests (determine viurs load in sample)
  • biological test (determine the number of specific mRNA molecules in a sample)
  • Principle: rely on detection and quantification of a fluorescent signal which is proportional to the amount of formed PCR product
Lösung ausblenden
TESTE DEIN WISSEN

PCR Components?

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TESTE DEIN WISSEN
  • Template
  • 2 Primers per template
  • Buffer (buffer substance, salt)
  • thermostable Polymerase (thermophillic bacteria - Thermus aquaticus, pyrococcus furiosus)
Lösung ausblenden
TESTE DEIN WISSEN

Regulation of DNA Replication?

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TESTE DEIN WISSEN
  • Initiation of replication requires cis-acting elements: Origin of Replication (ORI)
  • preferentially A-T rich sequences - facilitated DNA helix opening
  • Initiator Proteins associate with ORI
  • promotes interaction with DNA Helicase
  • DNA Primase adds RNA primer
  • replication machinery starts with DNA replication in both directions
  • replication fork ist established
Lösung ausblenden
TESTE DEIN WISSEN

Usage of primers of cDNA

Lösung anzeigen
TESTE DEIN WISSEN
  • Oligo dT
    • + primes on poly A+mRNA
    • + crude RNA preps
    • -Rere RNA can be underpresented
  • Random pdN6
    • +large cDNA library
    • - cDNA can be small in size
    • - poly A- template can also be reverse transcribed
  • Gene specific
    • + cDNa library is enriched for specific mRNA
    • cDNA libraried can be made for specific applications, cloning all antibody encoding
Lösung ausblenden
TESTE DEIN WISSEN

Gel Electrophoresis?

Lösung anzeigen
TESTE DEIN WISSEN
  • Gene materials:
    • argarose gel (routine application, longer than 200 bp)
    • polyacrylamide (short DNA fragments
  • DNA wanders to anode because of phosphate groups
  • sieve effect letzs long DNA fragments migrate more slowly
Lösung ausblenden
TESTE DEIN WISSEN

RNA Polymerase?

Lösung anzeigen
TESTE DEIN WISSEN
  • migrates along DNA double helix
  • base information of DNA template strand is scanned in 3-5 direction
  • substrate: Ribonucleoside Triphosphate (ATP, UTP, ...)
Lösung ausblenden
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Q:

Watson and Crick?

A:
  • the composition of DNA varies from one species to another
  • total amounts of pyrimidine equal to the amounts of purine: A+G=C+T
  • after Frnklin and Gosline made a xray diffraction analysis they comined the information
    • Nucleotides from a linear chein, linked by phospodiester bonds
    • DNA froms a holical double strand
    • antiparallel organization
    • minor and major groove
    • 1 double helix turn - 10 bp
Q:

Identification of recombinant clones (electroporation)?

A:
  • after electroporation
    • many Ecoli will have taken up one plasmid
    • very few will contain 2 oder more
    • many Ecoli cells without plasmid DNA are present
  • each plasmid molecule carries the selection marker gene (pET11a -> ampicillin)
  • on a medium containing ampicillin only Ecoli that carry a plasmid are able to grow
Q:

Human genome size?

A:

3234 Mbp

Q:

Histone Code?

A:

numerous possible combinations

  • various effects on gene expression
  • highly dynamic and reversible
  • interaction between chromatin remodelling, modification histone variants -> impacts on gene expression


Q:

PCR History

A:
  • first proposed by Khorana in 1970
  • made public by Kary Mullis in 1985
  • method of the year in 1989
  • nobel prize in 1993
Mehr Karteikarten anzeigen
Q:

DNA sequencing?

A:
  • 2 types of DNA sequencing
    • Dideoxy cycle DNA sequencing:
      • for sequencing single clones
    • High troughput, next generation DNA sequencing:
      • for genome and transcriptome sequencing
Q:

qPCR?

A:
  • Aim: detection and quantitation of specific DNA in samples
  • medical tests (determine viurs load in sample)
  • biological test (determine the number of specific mRNA molecules in a sample)
  • Principle: rely on detection and quantification of a fluorescent signal which is proportional to the amount of formed PCR product
Q:

PCR Components?

A:
  • Template
  • 2 Primers per template
  • Buffer (buffer substance, salt)
  • thermostable Polymerase (thermophillic bacteria - Thermus aquaticus, pyrococcus furiosus)
Q:

Regulation of DNA Replication?

A:
  • Initiation of replication requires cis-acting elements: Origin of Replication (ORI)
  • preferentially A-T rich sequences - facilitated DNA helix opening
  • Initiator Proteins associate with ORI
  • promotes interaction with DNA Helicase
  • DNA Primase adds RNA primer
  • replication machinery starts with DNA replication in both directions
  • replication fork ist established
Q:

Usage of primers of cDNA

A:
  • Oligo dT
    • + primes on poly A+mRNA
    • + crude RNA preps
    • -Rere RNA can be underpresented
  • Random pdN6
    • +large cDNA library
    • - cDNA can be small in size
    • - poly A- template can also be reverse transcribed
  • Gene specific
    • + cDNa library is enriched for specific mRNA
    • cDNA libraried can be made for specific applications, cloning all antibody encoding
Q:

Gel Electrophoresis?

A:
  • Gene materials:
    • argarose gel (routine application, longer than 200 bp)
    • polyacrylamide (short DNA fragments
  • DNA wanders to anode because of phosphate groups
  • sieve effect letzs long DNA fragments migrate more slowly
Q:

RNA Polymerase?

A:
  • migrates along DNA double helix
  • base information of DNA template strand is scanned in 3-5 direction
  • substrate: Ribonucleoside Triphosphate (ATP, UTP, ...)
Introduction to Molecular Biology

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