Archive

  • Visit JGI.DOE.GOV
Archived Educator Resources
Home › Archived Educator Resources › Sanger Sequencing Archive › How Sanger Sequencing Was Done › Step 5: Colony Picking

Step 5: Colony Picking

A 384-well culture stock plate.

A 384-well culture stock plate.

Believe it or not, millions of copies of the DNA fragments are just not enough; we need billions! So each colony needs to be cultured again. This entails picking the good colonies from the agar plates and transferring each one to a well on a 384-well plate.

Instead of taking a chance that a technician might go blind picking colonies all day, we use a robot called the Genetix Q-Pix. JGI has four pickers that operate 24 hours a day. The robots’ imaging system can discriminate between white and blue colonies. Each colony is put into a separate well that contains 70 µl of a nutrient-rich liquid medium.

A technician loads plates into the colony picking machine.

A technician loads plates into the colony picking machine.

All together, JGI can produce 500 384-well plates in a day (but we average 240). That’s 192,000 colonies! The plates are incubated for 18-20 hours at 37°C. After incubating, they can be stored at -80°C until they are ready for production sequencing.

  • How Sanger Sequencing Was Done
    • Step 1: Shearing of DNA
    • Step 2: Insertion of Fragments into a Plasmid
    • Step 3: Transformation
    • Step 4: Sub-Cloning the Sheared Fragment
    • Step 5: Colony Picking
    • Step 6: Lysing the Cell
    • Step 7: Rolling-Circle Amplification
    • Step 8: Sequencing Chemistry
    • Step 9: Post-Sequencing-Reaction Cleanup
    • Step 10: Capillary Sequencing
    • Step 11: Assembly
    • Step 12: Quality Assessment

More from the JGI archives:

  • Software Tools
  • Science Highlights
  • News Releases
  • Blog
  • User Proposals
  • 2018-24 Strategic Plan
  • Progress Reports
  • Historical Primers
  • Legacy Projects
  • Past Events
  • JGI.DOE.GOV
  • Disclaimer
  • Accessibility / Section 508
Lawrence Berkeley National Lab Biosciences Area
A project of the US Department of Energy, Office of Science

JGI is a DOE Office of Science User Facility managed by Lawrence Berkeley National Laboratory

© 1997-2025 The Regents of the University of California