Lecture #49

  1. REal time PCR
    • you have a fluorescence probe
    • you can see the amplication with each PCR cycle 
    • exponentially amplified DNA
  2. PCR applications
    • Genetic variations
    • Detection of rare sequences
  3. Application of digital PCR
    • rare  detetion 
    • residual disease in patients recieving chemotherapy and/or hematopietic cell transplantation for hematologic malignancies
  4. PCR forensics
    • can detect DNA fingerprinting
    • by looking at mother and father chromosomes
  5. Microarrays
    • take sample MRNA from two sample and mix them them with different courescent die. 
    • you hybridize them to the microarray 
    • sus normal cells 

    you can see how expressions differ based on color
  6. advantages of microarrays
    • automated and throughput
    • less hands on
  7. flow cytometry
    • asses size granularilty and protein expression of indiviual cells samples 
    • flourescently label a antibody that attached to a cell then they are detected which cells they bind to
    • you can see which kind of cells are expressing specific cellular molecules
  8. Elisa technique
    • enzyme inked immunosorbent assay 
    • direct - a plate with antigen bond to the plate. upon biniding it will emit a callot
    • indirect- a primary and secondary antibody that can bind to a subtrate thatgives off a certain color when binding occurs
  9. southern blotting
    dna is seperated based on size ( base pairs)
  10. technique o
  11. f
  12. sout
  13. blo
  14. tti
  15. technique of southern blotting
    • get DNA sample
    • run gel electrophoresis
    • put a nitrocellulaer membrane on the fell to transfer the DNa 
    • transfer it for radioactive development 
    • then finally detect
  16. northern blot application
    RNA detection
  17. SPEP
    • serum protein electrophoresis 
    • comassia staining is used
  18. what are the 2 major types of DNA sequencing
    • class chain termination Sanger sequencing ( first generation sequencing)
    • next generation sequencing
  19. mechanism of Sanger Sequencing
    • use ddNTop( dideoxynucleutide) and normal dNTPs
    • this will create random ending chains of Dna 
    • you will get each dna ending at every single nucleotide and you will be able to tell the sequence
  20. mec
  21. technique beh
  22. technique behind Next generation sequencing
  23. applications of New generation applications
  24. gene modification strategies
    • Gene therapy 
    • Gene KNock out/ knock in 
    • gene knock down
    • gene editing
  25. gene therapy technique
  26. gene therapy challeneges
    • coat
    • possibility of tumor
    • unwanted immune system reaction 
    • targeting wrong cells 
    • ....
  27. Gene knock out
    deleting a gene in a stem cell
  28. what is Gene knock down
    Rna induced silencing of a gene
  29. CRISPR/Cas9
    • cas9 and guide RNA 
    • cas9 deletes the unwanted base gene
Card Set
Lecture #49
Application of molecular Genetics II