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2. • Genetic markers resulting from PCR amplification of genomic DNA
segments recognized by random primers of arbitrary nucleotide
sequence.
• primers usually 10 mers
• GC rich.
• annealing temperature: low 35° C.
• amplified products are detected in agarose gels
3. PCR
• Buffer (containing Mg++)
• Template DNA
• 2 Primers that flank the fragment
of DNA to be Amplified
• dNTPs
• Taq DNA Polymerase (or another
thermally stable DNA polymerase)
RAPD
• Buffer (containing Mg++) usually
high Mg++ concentrations are used
lowering annealing stringency
• Template DNA
• 1 short primer (10 bases)not
known to anneal to any specific
part of the template DNA
• dNTPs
• Taq DNA Polymerase (or another
thermally stable DNA polymerase)
4.
5.
6. Advantageous
• Primer sequences are synthesized at
random (does not require any
sequences information).
• No gel transfer or hybridization is
required.
• Fast (many of the steps are
automated ).
• Relatively inexpensive
• Highly variable.
• The amount of template DNA is
extremely small.
Disadvantages
• Markers are dominant ( Presence of a
band could mean the individual is
either heterozygous or homozygous
for the sequence--can’t tell which ).