Summary: Personalized Medicine Alles

Read the summary and the most important questions on Personalized medicine alles

• 1 PCR

  This is a preview. There are 1 more flashcards available for chapter 1
  Show more cards here

• What is the goal of PCR?

  Exponential amplification of specific target piece of DNA

• How does is work? (3 steps)

  Monster -> DNA extractie
  You need 
  

  • DNA polymerase = taq polymerase
  • primers 
  • template DNA 
  • nucleotides

  
  

  1. Denaturation of the template strand into single strands
  2. Annealing of primers (of specific target DNA) to each original strand
  3. Extension of the new DNA strand from the primers with DNA-polymerase.

  
  

• 2 Sanger

  This is a preview. There are 2 more flashcards available for chapter 2
  Show more cards here

• What is the goal of sanger sequencing?

  Determine the precise order of the nucleotides of a given DNA fragment.

• What do you need for sanger seq?

  1. Primer 
     
  2. DNA template
  3. DNA polymerase 
  4. dNTP 
  5. ddNTP (=stop)

• How do you separate DNA fragments on length and charge?

  Gel electroforesis.
  

• What can you detect with sanger sequencing?What is an disadvantage of sanger?

  Sanger sequencing is the gold standard method for accurate detection of
  

  • single nucleotide variants
  • small insertions/deletions. 
  • Not so well duplications

  
  
  Disadvantage:
  - only short parts of DNA
  - Long parts => long time

• 3 NGS/wes

  This is a preview. There are 3 more flashcards available for chapter 3
  Show more cards here

• What is the goal of NGS?

  One gene or a gene package sequencen -> finding the order of the nucleotides.

• So what is the difference with sanger?

  • The length of the DNA sample: NGS is massively parallel, that is, sequencing millions of fragments simultaneously per run
  • NGS is better in detecting small variants and rare mutations 
  • Higher discovery power
  • Higher sensitivity 

• What is a problem with NGS/WES?

  You sequence short elements => can lead to unneccecairy frame shift. 
  You need to realine the different parts => difficult with copy number variants (CNV).  

• 3.1 Variant analysis

  This is a preview. There are 3 more flashcards available for chapter 3.1
  Show more cards here

• So you found variant/mutations. What is next? What do you want to know?

  • Is there a gene-disease association
  • Does this explain the pathogenic mechanism of the disease
  • Inheritance: autosomal dominant, recessive, x-linked
  • Prevalence, penetrance