One-Step Bacterial Artificial Chromosome (BAC) Modification: Cloning of the A Homology Arm into Reporter-Shuttle Vector.
Journal
Cold Spring Harbor protocols
ISSN: 1559-6095
Titre abrégé: Cold Spring Harb Protoc
Pays: United States
ID NLM: 101524530
Informations de publication
Date de publication:
01 07 2020
01 07 2020
Historique:
entrez:
3
7
2020
pubmed:
3
7
2020
medline:
8
6
2021
Statut:
epublish
Résumé
In this protocol, the homology arm sequence for one-step bacterial artificial chromosome (BAC) modification is introduced by ligation into the shuttle vector carrying the reporter sequence 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. To provide further assurance that the homology box has been successfully integrated into the plasmid, the enzyme digestion pattern of the modified plasmid is compared with that of the unmodified plasmid.
Identifiants
pubmed: 32611778
pii: 2020/7/pdb.prot098111
doi: 10.1101/pdb.prot098111
doi:
Substances chimiques
DNA, Bacterial
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
098111Informations de copyright
© 2020 Cold Spring Harbor Laboratory Press.