Education

Bioenergy at DOE JGI Introduction to Genomics How Sequencing Is Done Assembly Exercise Historical Timeline Tours Educational Links

How Sequencing Is Done


Animation of the JGI production sequencing process
(Flash, 30 minutes, 3.5 MB)

At JGI, we use whole-genome shotgun sequencing. This is a technique for determining the DNA sequence of a genome by randomly shearing the DNA, sequencing multiple fragments whose sequences overlap, and inferring the original sequence by reassembling the fragments. Three sizes of fragments are sequenced, 2-4 kb (kilobase, or 1000 bases), 8-10 kb, and 40 kb. This explanation follows the procedure for 2-4-kb fragments. Procedures for the other sizes are similar.

I.Library Creation

            1. Shearing of DNA

            2. Insertion of Fragments into a Plasmid

            3. Transformation

            4. Subcloning the Sheared Fragment

            5. Colony Picking

II.Production Sequencing

            6. Lysing the Cell

            7. Rolling-Circle Amplification

            8. Sequencing Chemistry

            9. Post-Sequencing-Reaction Cleanup

            10. Capillary Sequencing

III.Assembly and QA

            11. Assembly

            12. Quality Assessment