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DNA sequencing technology

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DNA sequencing technology. CMSC 828N, Fall 2008. Restriction enzymes. Library construction. First, do size selection run the DNA on a gel Cut out a swath of DNA for the desired size repeat [Switch to cloning slides here]. After cloning. Lyse (cut open and kill) E. coli with plasmids

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DNA sequencing technology

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  1. DNA sequencing technology CMSC 828N, Fall 2008

  2. Restriction enzymes

  3. Library construction • First, do size selection • run the DNA on a gel • Cut out a swath of DNA for the desired size • repeat • [Switch to cloning slides here]

  4. After cloning • Lyse (cut open and kill) E. coli with plasmids • Amplify sequences directly from built-in primers

  5. DNA sequencing

  6. Dideoxy terminators (modified DNA)

  7. Extend the sequences

  8. Run sequences through a gel

  9. Fluorescent dyes

  10. Animated gel separation

  11. Multiple “lanes” at one time

  12. Chromatogram data

  13. PCR • Amplify any DNA using 2 primers • Details here

  14. PCR animation

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