Molecular Biology
Lesson 3 of 4 10 min +90 XP

PCR & DNA Sequencing

Copying and reading the genetic code.

What you'll learn

  • Explain what PCR does
  • Describe the role of temperature cycling
  • State what DNA sequencing reveals
The molecular photocopier

To read a single sentence you first need many copies. PCR is a molecular photocopier that doubles a snippet of DNA again and again until there's enough to sequence — to spell out its letters and read the genetic message.

Amplify, then read

The polymerase chain reaction (PCR) makes millions of copies of a target DNA segment. Repeated cycles of heating and cooling denature the double helix, let short primers anneal to the flanking sequence, and let a heat-stable polymerase extend new strands — doubling the DNA each cycle. Sequencing then determines the exact order of bases (A, T, C, G).

Exponential copying

Each PCR cycle roughly doubles the target, so n cycles yield about 2ⁿ copies — 30 cycles turn one molecule into a billion, enough to detect and sequence.

PCR is the basis of many diagnostic tests: amplify a pathogen's DNA/RNA until it's detectable, even from a tiny sample.
Lab · Why cycle the temperature?
  1. Recall that DNA's two strands must separate before copying.
  2. Note that primers must then bind, and polymerase must extend.
  3. Explain why each step needs a different temperature.

What you should see: High heat denatures (separates) the strands, cooling lets primers anneal, and a mid temperature lets the polymerase extend — so cycling the temperature drives each stage of copying.

Knowledge Check

+25 XP / correct

1. The purpose of PCR is to…

2. Roughly how many copies result after 30 ideal PCR cycles from one molecule?