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Long Range DNA Polymerase, 2.5 U/µl


  • High-productivity, long-range PCR
  • Increased fidelity for accurate amplification of GC-rich templates
  • Polymerase mix for high yield and short cycle times


  • Long-range PCR up to 40 kb
  • Amplification of GC-rich templates

Purchase Long Range DNA Polymerase, 2.5 U/µl

Availability: In stock Bulk requests please inquire at

Catalog # Size Package Content Price* Qty
BR0300301 100 Units 40 µl Long Range PCR Enzyme Mix
400 µl 10X Long Range Reaction Buffer
1.5 ml 5X PCR Enhancer
BR0300302 500 Units 5 × 40 µl Long Range PCR Enzyme Mix
5 × 400 µl 10X Long Range Reaction Buffer
1.5 ml 5X PCR Enhancer

*Does not include applicable tax, shipping & handling, or other charges.


biotechrabbit™ Long Range DNA Polymerase is a first-choice for amplification of targets up to 40 kb in size with higher accuracy than Taq DNA polymerase.

This specially designed blend of thermophilic polymerases is well suited for amplification of targets that are GC-rich and have complex structures.

For the most demanding applications, the supplied 5× PCR Enhancer can be optionally used to improve results when using templates with GC-rich sequences and complex structures.

Long Range DNA Polymerase produces a mixture of A-tailed and blunt-end PCR products. It is advisable to blunt products before cloning into blunt-end vectors.




Long Range PCR Enzyme Mix

Long Range PCR Enzyme Mix, 2.5 U/µl, in storage buffer containing 50% (v/v) glycerol

10× Long Range Reaction Buffer

Optimized PCR reaction buffer including magnesium ions

5× PCR Enhancer

Proprietary PCR enhancer mix



−20°C (until expiry date – see product label)


Unit Definition

One unit catalyzes the incorporation of 10 nmol of deoxyribonucleotides into a polynucleotide fraction in 30 minutes at 72°C.

Quality Control
Functional assay

Human genomic DNA was amplified using the DNA Polymerase and specific primers to produce a distinct band of 750 bp.

Self-priming activity

Standard PCR is carried out without primers, using the DNA Polymerase and human genomic DNA. No products were amplified.

Exonuclease assay

Linearized lambda/HindII fragments are incubated with the DNA Polymerase in a 50 µl reaction mixture for 4 h at 37°C. No degradation of DNA was observed.

Endonuclease assay

lambda DNA is incubated with the DNA Polymerase in a 50 µl reaction mixture for 4 h at 37°C. No degradation of DNA was observed.

Nick Activity

Supercoiled plasmid DNA is incubated with the DNA Polymerase in a 50 µl reaction mixture for 4 h at 37°C. No conversion of covalently closed circular DNA to nicked DNA was detected.

E. coli DNA contamination assay

A sample of the denatured DNA Polymerase is analyzed with specific primers targeting the 16S rRNA gene in qPCR for the presence of contaminating E. coli DNA. No E. coli DNA was detectable.


Prevention of PCR contamination

When assembling the amplification reactions, care should be taken to eliminate the possibility of contamination with undesired DNA.

  • Use separate clean areas for preparation of samples and reaction mixtures and for cycling.
  • Wear fresh gloves. Use sterile tubes and pipette tips with aerosol filters for PCR setup.
  • Use only water and reagents that are free of DNA and nucleases.
  • With every PCR setup, perform a contamination control reaction that does not include template DNA.
Standard PCR setup

The standard PCR protocol using biotechrabbit reaction buffer provides excellent results for most applications. Optimization might be necessary for certain conditions, such as the amplification of long targets, high GC or AT content, strong template secondary structures or insufficient template purity. In such cases, optimization of template purification (see biotechrabbit nucleic acid purification kits), primer design and annealing temperature is recommended.

The best conditions for each primer-template can be optimized with the following:

  • Choosing the optimal quantities of template and primers
  • Determining optimal concentrations of the enzyme and magnesium ions
  • Optimizing cycling conditions

If unspecific amplification occurs, the amount of DNA Polymerase and the primer concentration can be reduced. Correspondingly, these can be increased when yield is low.

Basic Protocol
  • Optionally, use the supplied 5× PCR Enhancer to increase the yield and to lower the background in more complicated PCR reactions (low amounts of template, impure or GC-rich template).
  • Thaw on ice and mix all reagents well, especially the MgCl2 solution and dNTPs.
  • Keep all reagents and reactions on ice.
  • When setting up multiple reactions, prepare a master mix of water, buffer, dNTPs and polymerase. Prepare enough master mix for one more than the actual number reactions. Alternatively, use biotechrabbit Long Range PCR Master Mix, 2× (cat. no. BR0300401)
  • Pipet the master mix into thin-walled 0.2 ml PCR tubes.
  • Add template and primers separately if they are not used in all reactions.



Final concentration

10× Long Range Reaction Buffer

5 µl

5× PCR Enhancer (optional - BR1900201)

10 µl

10 mM dNTP Mix

1 µl

200 µM

Forward primer


0.2–1 µM

Reverse primer



Template DNA


10 pg–1 μg


Use 0.01–1 ng for plasmid or phage DNA and 0.1–1 μg for genomic DNA

Long Range PCR Enzyme Mix (2.5 U/µl)

0.5–1 µl

1.25–2.5 U

Nuclease free water



Total volume

50 µl


  • Mix and centrifuge briefly to collect the liquid in the bottom of the tube.
  • Place in the PCR cycler.
Cycling Program





Initial activation


2 min




30 s




30–45 s



Approximately 5°C below Tm of primers



30 s/kb


For fragments longer than 5 kb, use 68°C extension temperature and 1 min/kb timing.

Final extension


5 min



To extend all incomplete PCR products

Storage in the cycler





  • Add loading dye solution (see DNA Loading Dye, 6×, cat. no. BR0800301) to the reactions to analyze PCR products on a gel or store them at −20°C.
  • For cloning, always purify the PCR product from a gel (see BR0700401 GenUP™ Gel Extraction Kit).
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