Lecture 10 - Extra Flashcards
What are SNPs?
Single nucleotide polymorphism
A change in 1 nucleotide base in a DNA sequence
What is a nucleotide base?
Adenine Thymine
Guanine Cytosine
What are the effects of SNPs?
Physiology Personality Racial/Geographical differences Disease susceptibility Cancer susceptibility Radiation susceptibility
How are single nucleotide polymorphisms (SNPs) detected?
Maldi-ToF
Genotyping using PCR amplification
Primer extension
MS detection
What is small molecule quantitation?
SRM-MS used in clinical diagnosis/pharmaceuticals
Drug testing
3 variations:
Standard addition experiment
-MS response relative to drugs consumed
External and internal standard
-Biological sample > Internal standard > Purification > External standard > MS analysis
_______
Internal standard is best method as losses in purification and MS performance are accounted for
What are FDA requirements for new proteomic methods?
Selectivity Sensitivity Linearity Recovery Accuracy Sample stability
What reaction does phenylalanine hydroxylase take part it?
Phenylalanine -> Tyrosine
PA build up causes brain damage, seizures, mental retardation
What is metabolomics?
Profiling metabolites without bias towards a specific metabolite or group or metabolites
Characterisation of metabolites
Define metabolomics
Quantification of the total metabolites complement inside and outside a cell under defined conditions
Define exometabolome
Total metabolites excreted outside the cell (culture supernantant)
Define endometabolome
Total metabolites located inside the cell (intracellular)
Define footprinting
Qualitative analysis of exometabolome
e.g. metabolites excreted by kidneys
Define fingerprinting
Qualitative analysis of endometabolome
e.g. herbal medicine analysis, no purity information
Define target analysis
Quantitative analysis of known pre-defined metabolites
hypothesis driven
Define quenching
Instantaneous arrest of endogenous metabolic activity
