Principles of PCR

1
Principles of PCR

• MUPGRET Workshop
• June 18, 2004

2
PCR

• Polymerase Chain Reaction
• http//www.dnai.org/b/index.html
• http//palou.uib.es/roolpi/PCR/

3
Basic PCR

• Initially reagents in excess, template and
  product dont renature and outcompete primer
  binding, so amplification occurs at an
  exponential rate.
• Timepoint when exponential turns to linear rate
  is different depending on the initial
  concentration of the target molecule.

4
Basic PCR

• To properly quantify one product relative to
  another you need to be in the exponential phase
  of amplification for both and know when the end
  point occurs.

5
Troubleshooting PCR

• http//www.promega.com/guides/pcr_guide/070_22/pro
  mega.html
• http//info.med.yale.edu/genetics/ward/tavi/Trbles
  ht.html
• http//palou.uib.es/roolpi/PCR/

6
Electrophoresis

• A way to separate DNA molecules.
• http//gslc.genetics.utah.edu/units/biotech/gel/