Bioanalytik at Universität Tübingen | Flashcards & Summaries

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Lernmaterialien für bioanalytik an der Universität Tübingen

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Database repertoires

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  1. Sequence data base (ex. NCBI)
    • DB for DNA/RNA sequences
  2. Database for other primary data:
    • expression data, metabolome data, protein crystal structure
  3. secondary DB (ex. EXPASY):
    • contain info derived from primary DB
  4. cumulative DB (ex. BRENDA):
    • collect data bout specific subject/organism
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Electrochemical detection (EC)

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1. Pulsed Amperometry: 

- Voltage between two electrodes.

- Voltage changes if analyte is oxidized at anode or reduced at cathode

-Destructive and electrodes might degrade 


2. Condictivity Detection:

- Constant potential between two electrodes

- Passing analytes change the conductivity

- Non-destructive but less sensitivity

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Gases

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  • mostly easy sample taking & relatively clean sample
  • Gas mouse 
  • liquids that absorb gases
  • integrated sampling with solid absorbing materials
  • small gas molecules can diffuse trough container !
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UV/Vis Detector

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  • measures light absorption at predefined wavelength (190-1000 nm)
  • Prism splits light from source 
  • slit blocks all but selected wavelength
  • versatile
  • non-destructive
  • photomultiplier detector
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FSC

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  • Forward scattered light
  • info on size of particle 
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Sample taking

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  • depends on matrix
  • do not affect sample during sample taking
  • cleanliness of containers and labs essential
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Ion Exchange (IEX) Chromatography

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Stationary phase: 

  • Anion exchanger: positively charged sp binds anions
  • Cation exchanger: neg charged sp binds cations

Mobile phase:

  • Buffer 
  • Consider: exchanger & analyte MUST be charged; select good buffer for right pH
  • High capacity but low separation power 
  • Used for proteins & DNA
  • EXCELS at Amino acid separation
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What is retention time tR

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time for analyte to reach detector

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Column types

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  • Packed: column filled with small particles that have pores
  • monolithic: no individual particles inside but 1 block with many gaps & pores
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Particle material

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Responsible for stability of column (solvents; pH; pressure)

  • Silica: stable at acidic pH not at basic; pressure stable
  • Polystyrol-, Acrylate-polymers: wide pH stability; low pressure stability
  • Sugar polymers: low pH and low pressure stable; biocompatible (hydrophilic); mainly for SEC
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Applications of FACS  in plant biology 

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  • DNA analysis
  • sorting of nuclei or protoplasts for transcriptome cell profiling 
  • gene-expression reporter assay 
  • quantification of transfection efficiency
Lösung ausblenden
TESTE DEIN WISSEN

Hydrophobic interaction chromatography (HIC)

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TESTE DEIN WISSEN
  • SP: non polar (aromatic surface)
  • MP: buffer; inverse salt gradient from strong to weak 
  • Hydrophobic & aromatic interactions
  • For when RP/ NP fail PROTEINS; NUCLEIC ACIDS
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  • 103 Lernmaterialien

Beispielhafte Karteikarten für deinen bioanalytik Kurs an der Universität Tübingen - von Kommilitonen auf StudySmarter erstellt!

Q:

Database repertoires

A:
  1. Sequence data base (ex. NCBI)
    • DB for DNA/RNA sequences
  2. Database for other primary data:
    • expression data, metabolome data, protein crystal structure
  3. secondary DB (ex. EXPASY):
    • contain info derived from primary DB
  4. cumulative DB (ex. BRENDA):
    • collect data bout specific subject/organism
Q:

Electrochemical detection (EC)

A:

1. Pulsed Amperometry: 

- Voltage between two electrodes.

- Voltage changes if analyte is oxidized at anode or reduced at cathode

-Destructive and electrodes might degrade 


2. Condictivity Detection:

- Constant potential between two electrodes

- Passing analytes change the conductivity

- Non-destructive but less sensitivity

Q:

Gases

A:
  • mostly easy sample taking & relatively clean sample
  • Gas mouse 
  • liquids that absorb gases
  • integrated sampling with solid absorbing materials
  • small gas molecules can diffuse trough container !
Q:

UV/Vis Detector

A:
  • measures light absorption at predefined wavelength (190-1000 nm)
  • Prism splits light from source 
  • slit blocks all but selected wavelength
  • versatile
  • non-destructive
  • photomultiplier detector
Q:

FSC

A:
  • Forward scattered light
  • info on size of particle 
Mehr Karteikarten anzeigen
Q:

Sample taking

A:
  • depends on matrix
  • do not affect sample during sample taking
  • cleanliness of containers and labs essential
Q:

Ion Exchange (IEX) Chromatography

A:

Stationary phase: 

  • Anion exchanger: positively charged sp binds anions
  • Cation exchanger: neg charged sp binds cations

Mobile phase:

  • Buffer 
  • Consider: exchanger & analyte MUST be charged; select good buffer for right pH
  • High capacity but low separation power 
  • Used for proteins & DNA
  • EXCELS at Amino acid separation
Q:

What is retention time tR

A:

time for analyte to reach detector

Q:

Column types

A:
  • Packed: column filled with small particles that have pores
  • monolithic: no individual particles inside but 1 block with many gaps & pores
Q:

Particle material

A:

Responsible for stability of column (solvents; pH; pressure)

  • Silica: stable at acidic pH not at basic; pressure stable
  • Polystyrol-, Acrylate-polymers: wide pH stability; low pressure stability
  • Sugar polymers: low pH and low pressure stable; biocompatible (hydrophilic); mainly for SEC
Q:

Applications of FACS  in plant biology 

A:
  • DNA analysis
  • sorting of nuclei or protoplasts for transcriptome cell profiling 
  • gene-expression reporter assay 
  • quantification of transfection efficiency
Q:

Hydrophobic interaction chromatography (HIC)

A:
  • SP: non polar (aromatic surface)
  • MP: buffer; inverse salt gradient from strong to weak 
  • Hydrophobic & aromatic interactions
  • For when RP/ NP fail PROTEINS; NUCLEIC ACIDS
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