|Title||Two-Step Bacterial Artificial Chromosome (BAC) Engineering: Cloning of the A and B Homology Arms into the Shuttle Vector.|
|Publication Type||Journal Article|
|Year of Publication||2020|
|Authors||Heintz N, Gong S|
|Journal||Cold Spring Harb Protoc|
|Date Published||2020 04 01|
This protocol describes the preparation of the shuttle vector before its introduction into bacterial artificial chromosome (BAC) host cells for BAC two-step engineering. The homology arm sequences, prepared previously, are introduced by ligation into the digested shuttle vector DNA to provide sites for recombination within the BAC clone. Crude lysates of individual bacterial transformants serve as templates in polymerase chain reaction (PCR) analysis to confirm the presence of the homology arms in the recombinant shuttle vector.
|Alternate Journal||Cold Spring Harb Protoc|