Polymerase Chain Reaction PCR 1

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• Polymerase Chain Reaction (PCR) 1

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Polymerase Chain Reaction (PCR) 2
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PCR of mRNA
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Semi quantitative Reverse transcriptase PCR
(RT-PCR)
End-point reaction after 40 cycles of
replication Agarose gel with ethidium bromide dye
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End point analysis
12 cycles
25 cycles
40 cycles
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Problems of quantitative end-point PCR

• Variation of end- point even with identical
  samples
• Less variation in midpoint of reaction

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Quantitative Real time PCR
Threshold
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SYBR Green fluorophore
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Polymerase Chain Reaction (PCR) 3 SYBR Green
fluorescence measure after every cycle