Whole-genome shotgun sequencing is a technique used 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.
Library Creation
2. Insertion of Fragments into a Plasmid
4. Sub-cloning the Sheared Fragment
Production Sequencing
7. Rolling-Circle Amplification
9. Post-Sequencing-Reaction Cleanup
Assembly and QA