06 PCR
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PCR
- DNA polymerase requires primer
- used for small amount of DNA curculating
- HIV: RNA. Use reverse transcriptase to make copy of virus RNA: cDNA
Blotting
- put a piece of filter paper on top. DNA stick to it
- DNA left complementary to probe
- for characterizing genes
- length of fragments unique to each particular gene
- Restriction nuclease added to Gene A and Gene B: breaking up into different fragments
- Compare unknown gene that break up to gene A or B
- same technique for RNA
- studies gene expression by finding mRNA corresponding to standard
- use antibody to protein instead of probe
Flow Cytometry
- side and forward scatter
- forward scatter based on cell size
- side scatter based on granularity
- lymphocytes: small, little granules
- granulocytes: big, lots of granules
- determine Hb F in maternal circulation = hemorrhage
Elisa
- add serum to one of the wells
- plate designed so antigen stick to surface
- wash away serum and left with antigens bound
- direct: for every antigen test for, must have an antibody enzyme linked
- indirect: more common
- unbound antibody: free antibody from last slide
- any HIV antibody in sample will bind to antigen in plate
Microassays
FISH
- like microarray, uses fluorescent
- tell that gene is present on that chromosome
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