06 PCR

PCR

• DNA polymerase requires primer

• variant, quantitative

• used for small amount of DNA curculating

• HIV: RNA. Use reverse transcriptase to make copy of virus RNA: cDNA

Blotting

• binding = hybridization

• 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

• determine genotype

• 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

• good for detect proteins

• 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