QRT-PCR/Single tube

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This protocol describes one-step real time quantitivie PCR to quantify relative levels of RNA in

Starting Materials

  • Validated PCR primers that are efficient over the range of RNA that you are assaying.

Basic Principle

  • RNA is reverse transcribed in a single well.
  • PCR reaction procedes. The levels of template are quantified during each cycle.

Protocol

Analysis

Delta-Delta Ct Method. See Livak KJ, Schmittgen. Methods 25 402-408 (2001)