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Lecture 6: Probes, DNA amplification and genomic libraries - Coggle Diagram

- - - - above Tm, will inactivate and primers wont bind
      - solution?
        
        Thermostable polymerase - >20 cycles
        
        Temperature cycling
  - - - eg. if primer contains a change, site specific mutation occurs
    - - breast cancer
        
        BRCA1 and BRCA2 produce
        tumor suppressor proteins
        
        BRCA proteins not present - maybe cancer
      - Paternity testing
        
        short tandem repeat (STR)
        
        Children inherit half their alleles from each parent – should possess a combination of parents alleles
      - Forensic Science
        
        Variable Number Tandem Repeats (VNTR) - highly variable in population
        
        longer tha STR (100bp)
        
        inherit each VNTR locus from their
        mother and father
        
        A pair of primers that bracket this VNTR loci will provide bands of different sizes as inherited from each parent
        
        Two unrelated individuals usually do not share same pair of sequences
        
        use gel electrophoresis
- - - - 1/2 for each E. Coli cell
      - preferred vector
  - - - not representative of transcriptome
- - - - primer - Poly T tail hybridise w Poly A tail
      - RNase H makes nicks and gaps in RNA
      - DNA polymerase copies single stranded DNA into double stranded cDNA
        
        RNA used as primers
- - - - attachment of probes prepared separately
    - - in situ synthesis of oligonucleotide probes