TA CloningTA Cloning uses the terminal transferase activity of some DNA polymerases. In TA Cloning Taq polymerase adds a 3'-A overhang to each end of the PCR product. TA Cloning makes it possible to clone the PCR product into a cloning vector with 3'-T overhangs. TA Cloning is widely used in molecular biology.
Edited by: Manuel Fuentes and Joshua LaBaer"many excellent chapters" (Doodys) read more ...
An overview of targeted proteomics in biomedical science.
TA CloningTA Cloning exploits the terminal transferase activity of some DNA polymerases. In TA Cloning Taq polymerase adds a 3'-A overhang to each end of the PCR product. TA Cloning makes it possible to clone the PCR product into a cloning vector with 3'-T overhangs. During TA Cloning the PCR products with dA overhang are mixed in high proportion with the vector. The complementary overhangs of T vector and PCR product are then ligated using T4 DNA ligase. "TA cloning" is a popular method of cloning without the use of restriction enzymes; instead, PCR products are amplified with only Taq DNA polymerase and other polymerases. These polymerases lack 5'-3' proofreading activity and add an adenosine triphosphate residue to the 3' ends of the double-stranded PCR products. Such PCR amplified products can thus be cloned in a linearized vector that has complementary 3' thymidine triphosphate overhangs. A recently described Quick Assemble PCR cloning protocol has been used to improve TA cloning. The new 2-day PCR-based method can be used to generate both circularized and linear final assembled products. The protocol circumvents the use of DNA ligase. The PCR-based protocol uses a TA cloning vector as the template for the linear vector backbone demonstrating the ability to improve this technique. from Zuo and Rabie in Curr. Issues Mol. Biol. 12: 11-16
- Aquatic Biofilms
- Thermophilic Microorganisms
- Flow Cytometry in Microbiology
- Probiotics and Prebiotics
- Corynebacterium glutamicum
- Advanced Vaccine Research Methods for the Decade of Vaccines
- Bacteria-Plant Interactions
- Metagenomics of the Microbial Nitrogen Cycle
- Pathogenic Neisseria
- Human Pathogenic Fungi
- Applied RNAi
- Molecular Diagnostics
- Phage Therapy
- Bioinformatics and Data Analysis in Microbiology
- The Cell Biology of Cyanobacteria
- Pathogenic Escherichia coli
- Campylobacter Ecology and Evolution