The introduction of new laboratory exercises "TA cloning and transformation of bacteria" in the teaching laboratories C653
Provider: Ministerstvo školství, mládeže a tělovýchovy
Programme: Interní rozvojová soutěž
Implementation period: 01.01.14 - 31.12.14
Workplace:
Fakulta chemicko-technologická - Katedra biologických a biochemických věd
Investigator: Pejchalová Marcela
Description:
Enzyme Taq polymerase used for PCR adds one deoxyadenosine at the 3'end of the amplicon. By TA cloning ligation of PCR products into the linearized vector with the cohesion ends (deoxythimidin at the 3'end). Ligation can be performed easily using the TA Cloning Kit. The ligation mixture was transformed into competent cells of Escherichia coli, which were transfered in a suitable selection medium. From bacterial colonies are isolated plasmid DNA and by agarose electrophoresis is controlled the integration into vector.
Enzyme Taq polymerase used for PCR adds one deoxyadenosine at the 3'end of the amplicon. By TA cloning ligation of PCR products into the linearized vector with the cohesion ends (deoxythimidin at the 3'end). Ligation can be performed easily using the TA Cloning Kit. The ligation mixture was transformed into competent cells of Escherichia coli, which were transfered in a suitable selection medium. From bacterial colonies are isolated plasmid DNA and by agarose electrophoresis is controlled the integration into vector.
