TaqStart Antibody
TaqStart Antibody
Brand: Takara Bio.
In stock
SKU
TaqStart Antibody
TaqStart Antibody: turn any Taq into a hot-start Taq
TaqStart Antibody provides antibody-mediated hot start that enhances the specificity and sensitivity of PCR. This antibody inhibits polymerase activity before the onset of thermal cycling, preventing nonspecific amplification and primer-dimer formation. When the reaction temperature is raised, the antibody is quickly inactivated and PCR proceeds. Dilution buffer is provided.
TaqStart Antibody is significantly more convenient to use than other hot-start methods and offers several advantages:
- Avoids sample damage due to depurination that can occur with the high-temperature incubations that are necessary to activate some hot-start enzymes (Fromenty et al. 2000; Barnes 1994; Fromenty et al. 1996; Friedberg, Walker, and Siede 1995; Lindahl and Andersson 1972; Suzuki, Ohsumi, and Makino 1994).
- Reduces the risk of cross-contamination because it is not necessary to reopen the reaction tubes after heating.
- Can be used when other hot-start methods are difficult to perform, such as for high-throughput PCR, in situ PCR, microtiter-plate formats, capillary PCR, and oil or wax-free environments.
- Provides more definitive PCR results in cases when amplification of nonspecific products is a problem, such as reactions involving low copy number targets, complex DNA background, or degenerate primers.
- TaqStart Antibody is effective with any Taq-derived DNA polymerase (native, recombinant, and N-terminal deletion mutants). Our Titanium Taq DNA Polymerase and all of our Advantage HD and Advantage 2 products include TaqStart Antibody.
Overview
- Allows convenient room temperature reaction setup
- Provides increased specificity, which leads to reduced background, allowing you to obtain your desired amplification product the first time
- Effective and inexpensive method for hot-start PCR
- Antibody can be used with any full-length Taq polymerase
- Available in bulk quantities
Applications
- Hot-start PCR
Increased yield and specificity of RT-PCR with TaqStart Antibody
Increased yield and specificity of RT-PCR with TaqStart Antibody. cDNA templates were reverse transcribed using oligo(dT)18 primers and 2 μg of total RNA isolated from human PHA-stimulated peripheral blood leukocytes or mouse RAW 264.7 macrophage cells. Taq DNA Polymerase was added either with (+) or without (–) TaqStart Antibody for 35 cycles. PCR products were run on a 1.8% agarose/EtBr gel. Lane 1: human c-jun; 410 bp. Lane 2: mouse IL-2 receptor; 700 bp. Lane 3: mouse inducible nitric oxide synthase; 497 bp. Lane M: FX174/Hae III DNA size marker.
TaqStart Antibody inhibits polymerase activity before thermal cycling begins
TaqStart Antibody inhibits polymerase activity before thermal cycling begins.
References
Barnes, W. M. PCR amplification of up to 35-kb DNA with high fidelity and high yield from lambda bacteriophage templates. Proc. Natl. Acad. Sci. U. S. A. 91, 2216–20 (1994).
Friedberg, E. C., Walker, G. C. & Siede, W. DNA repair and mutagenesis (ASM Press, 1995).
Fromenty, B. et al. Most cases of medium-chain acyl-CoA dehydrogenase deficiency escape detection in France. Hum. Genet. 97, 367–8 (1996).
Fromenty, B., Demeilliers, C., Mansouri, A. & Pessayre, D. Escherichia coli exonuclease III enhances long PCR amplification of damaged DNA templates. Nucleic Acids Res. 28, E50 (2000).
Lindahl, T. & Andersson, A. Rate of chain breakage at apurinic sites in double-stranded deoxyribonucleic acid. Biochemistry 11, 3618–23 (1972).
Suzuki, T., Ohsumi, S. & Makino, K. Mechanistic studies on depurination and apurinic site chain breakage in oligodeoxyribonucleotides. Nucleic Acids Res. 22, 4997–5003 (1994).
Product citations
Gerard, G. F., D'Alessio, J. M. in Methods Mol. Biol. (Burrell, M. M.) 73–94 (Humana Press, 1993).
Kellogg, D. E. et al. TaqStart Antibody: "hot start" PCR facilitated by a neutralizing monoclonal antibody directed against Taq DNA polymerase. Biotechniques 16, 1134–7 (1994).
Write Your Own Review
Increased yield and specificity of RT-PCR with TaqStart Antibody
Increased yield and specificity of RT-PCR with TaqStart Antibody. cDNA templates were reverse transcribed using oligo(dT)18 primers and 2 μg of total RNA isolated from human PHA-stimulated peripheral blood leukocytes or mouse RAW 264.7 macrophage cells. Taq DNA Polymerase was added either with (+) or without (–) TaqStart Antibody for 35 cycles. PCR products were run on a 1.8% agarose/EtBr gel. Lane 1: human c-jun; 410 bp. Lane 2: mouse IL-2 receptor; 700 bp. Lane 3: mouse inducible nitric oxide synthase; 497 bp. Lane M: FX174/Hae III DNA size marker.
TaqStart Antibody inhibits polymerase activity before thermal cycling begins
TaqStart Antibody inhibits polymerase activity before thermal cycling begins.
