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Amplification of human genomic DNA by Takara LA Taq DNA polymerase LA Taq
long distance pcr Hot-start LA Taq
LA Taq with GC buffers LA Taq with GC buffers
Home › Learning centers › PCR › Technical notes › Long-range PCR with LA Taq

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Amplification of human genomic DNA by Takara LA Taq DNA polymerase LA Taq
long distance pcr Hot-start LA Taq
LA Taq with GC buffers LA Taq with GC buffers
Tech Note

Long-range PCR with Takara LA Taq DNA polymerase

  • Example 1: Amplification of E. coli products up to 38 kb in length
  • Example 2: Amplification of E. coli products up to 18 kb in length
  • Example 3: Long-range PCR using human genomic DNA
Introduction 38 kb E. coli DNA 18 kb E. coli DNA Human gDNA Conclusion

Introduction  

Takara LA Taq DNA polymerase (Cat. # RR002A, also available in hot-start and high GC-compatible versions) is optimized for long-range PCR. These experiments show examples of long-range PCR using Takara LA Taq Polymerase with E. coli or human genomic DNA as the template.

38 kb E. coli DNA  

Example 1: Amplification of E. coli DNA up to 38 kb in length

Methods

100 ng of E. coli genomic DNA was used as the template in a 50-µl reaction. Takara LA Taq DNA polymerase (Cat. # RR002A) was used for amplification using the recommended conditions. After PCR, 5 µl of the reaction mixture was used for electrophoresis on a 0.4% agarose gel.

Results

Peak calling and electropherograms of pre- and post-PCR size selection libraries.

Figure 1. Products (20–38 kb) amplified from E. coli genomic DNA using Takara LA Taq DNA polymerase. Amplified product sizes are 20 kb (Lane 2), 28 kb (Lane 3), 30 kb (Lane 4), and 38 kb (Lane 5). Molecular weight markers: Lane 1: HindIII-digested lambda DNA; Lane 6: High-molecular weight marker.

18 kb E. coli DNA  

Example 2: Amplification of E. coli DNA up to 18 kb in length

Methods

PCR was performed using the following conditions outlined below. After PCR, 5 µl of the reaction mixture was used for electrophoresis on a 0.4% agarose gel.

Peak calling and electropherograms of pre- and post-PCR size selection libraries.

Results

Peak calling and electropherograms of pre- and post-PCR size selection libraries.

Figure 2. Products (2–18 kb) amplified from E. coli genomic DNA using Takara LA Taq. Amplified product sizes are 2 kb (Lane 2), 4 kb (Lane 3), 6 kb (Lane 4), 8 kb (Lane 5), 10 kb (Lane 6), and 18 kb (Lane 7). Lanes 1 and 8 contain HindIII-digested lambda DNA as a molecular weight marker.

Human gDNA  

Example 3: Long-range PCR using human genomic DNA 

Methods

The TPA gene region and the β-globin cluster region were each amplified from human genomic DNA using Takara LA Taq DNA polymerase. The reactions were set up as indicated below:

ComponentVolumeFinal concentration
Human genomic DNA (500 ng/µl) 1 µl 500 ng per 50 µl reaction
10x LA PCR Buffer II (Mg2+ Plus) 5 µl 1X
dNTPs 8 µl 400 µM each
Sense Primer (20 pmol/µl) 0.5 µl 0.2 µM
Antisense Primer (20 pmol/µl) 0.5 µl 0.2 µM
Takara LA Taq 0.5 µl 2.5 U per 50 µl reaction
Sterile distilled water 34.5 µl  

After the 50-µl reactions were prepared, each reaction was overlayed with an equal volume of mineral oil. PCR was performed using the following conditions:

Peak calling and electropherograms of pre- and post-PCR size selection libraries.

After PCR, 5 µl of the reaction mixture was used for electrophoresis on a 0.4% agarose gel.

Results

ChIP-seq library complexity and reproducibility from total DNA from small Amounts of cells. DNA SMART ChIP-seq library overlap with ENCODE data.

Figure 3. Products amplified from human genomic DNA using Takara LA Taq. LA Taq was used to amplify 17.5-kb and 21.5-kb fragments of the beta-globin gene cluster region (Lanes 2 and 3, respectively) and a 27-kb portion of the TPA gene region (Lane 4). Molecular weight markers: Lane 1: HindIII-digested lambda DNA; Lane 5: High-molecular weight markers.

Conclusion  

Amplification of E. coli genomic targets up to 38 kb in length or human genomic targets up to 27 kb in length was achieved using Takara LA Taq DNA polymerase.

Learn more about Takara LA Taq DNA polymerase »

Related products

Cat. # Product Size Price License Quantity Details
RR002A TaKaRa LA Taq® DNA Polymerase (Mg2+ free buffer) 125 Units USD $158.00

Takara LA Taq is a mixture of Taq Polymerase with a proofreading polymerase. The mixture is optimized for amplification of long DNA templates (routinely 20 kb and up to 48 kb depending on template type) with high fidelity. This LA Taq polymerase mix is supplied with separate tubes of optimized buffer (Mg2+ free), MgCl2, and dNTPs.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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Amplification of lambda DNA fragments ranging from 0.5-35 kb using Takara LA Taq DNA polymerase.

Amplification of lambda DNA fragments ranging from 0.5-35 kb using Takara LA Taq DNA polymerase.

Amplification of lambda DNA fragments ranging from 0.5-35 kb using Takara LA Taq DNA polymerase. The LA Taq enzyme was able to generate high yields of product even for very long fragments (28 kb).

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Amplification of human genomic DNA with Takara LA Taq DNA Polymerase

Amplification of human genomic DNA with Takara LA Taq DNA Polymerase

Amplification of human genomic DNA with Takara LA Taq DNA Polymerase. Purified human genomic DNA (500 ng in a 50 µL reaction) was used as a template for PCR amplification of regions of the beta-globin locus and the TPA gene with Takara LA Taq polymerase. The sizes of the amplified products were 17.5 kb (lane 1), 21.5 kb (lane 2), and 27 kb (lane 3). Lane M contains High MW Markers (Fisher Scientific).

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RR002A: TaKaRa LA Taq DNA Polymerase (Mg2+ free buffer)

RR002A: TaKaRa LA Taq DNA Polymerase (Mg2+ free buffer)
RR002B TaKaRa LA Taq® DNA Polymerase (Mg2+ plus buffer) 1,000 Units USD $1183.00

Takara LA Taq is a mixture of Taq Polymerase with a proofreading polymerase. The mixture is optimized for amplification of long DNA templates (routinely 20 kb and up to 48 kb depending on template type) with high fidelity. This LA Taq polymerase mix is supplied with separate tubes of optimized buffer (Mg2+ plus) and dNTPs. Cat. # RR002B contains 4 of Cat. # RR002M. Please refer to Cat. # RR002M for complete product documentation and resources.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents You May Also Like Resources
RR002C TaKaRa LA Taq® DNA Polymerase (Mg2+ plus buffer) 3,000 Units USD $3456.00

Takara LA Taq is a mixture of Taq Polymerase with a proofreading polymerase. The mixture is optimized for amplification of long DNA templates (routinely 20 kb and up to 48 kb depending on template type) with high fidelity. This LA Taq polymerase mix is supplied with separate tubes of optimized buffer (Mg2+ plus) and dNTPs. Cat. # RR002C contains 12 of Cat. # RR002M. Please refer to Cat. # RR002M for complete product documentation and resources.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents You May Also Like Resources
RR002M TaKaRa LA Taq® DNA Polymerase (Mg2+ plus buffer) 250 Units USD $315.00

Takara LA Taq is a mixture of Taq Polymerase with a proofreading polymerase. The mixture is optimized for amplification of long DNA templates (routinely 20 kb and up to 48 kb depending on template type) with high fidelity. This LA Taq polymerase mix is supplied with separate tubes of optimized buffer (Mg2+ plus) and dNTPs.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components You May Also Like Image Data Resources

Back

RR002M: TaKaRa LA Taq DNA Polymerase (Mg2+ plus buffer)

RR002M: TaKaRa LA Taq DNA Polymerase (Mg2+ plus buffer)

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