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Expand Long Template PCR System

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Order information
Expand Long Template PCR System material number and pack size:
Material Number Pack Size
03321053103 custom fill
Will be supplied as "Expand LT PCR Sys. Enzymmix, Bulk". Unit of measure is "kU".
The enzyme is supplied without reaction buffer.
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Overview
Proofreading blend for accurate amplification of genomic DNA targets up to 20 kb using PCR.
Enzyme blend consisting of Taq DNA Polymerase and Tgo DNA Polymerase.
  • Amplify long templates.
    Generate PCR products 5 to 20 kb in length from complex genomic DNA using this optimized enzyme blend.
  • Achieve higher yields and fidelity.
    Three times higher fidelity with higher yield compared to Taq DNA Polymerase.
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Application
Use Expand Long Template PCR System for:
  • Routine amplification of DNA fragments up to 20 kb from all DNA
  • Amplification of DNA fragments up to 40 kb from λDNA
  • Labeling of PCR products with modified nucleotides (e.g., DIG-dUTP, biotin-dUTP, fluorescein-dUTP)
  • Combination with dUTP and Uracil-DNA Glycosylase for prevention of carryover contamination between PCR reactions
  • Manufacture of amplification mixtures for regulated applications (e.g., in vitro diagnostics, quality control), including validation
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Specification
Enzymes in the Expand Long Template PCR System were originally isolated from the thermophilic eubacteria Thermus aquaticus (Taq) BM and Thermococcus gorgonarius (Tgo), both expressed in E. coli.
Enzyme acivities:
Taq Polymerase: Highly processive 5'-3' DNA polymerase, double-strand specific 5'-3' exonuclease, no 3'-5' exonuclease activity
Tgo Polymerase: Highly processive 5'-3' DNA polymerase, double-strand specific 3'-5' exonuclease (also known as proofreading activity), no 5'-3' exonuclease activity
Temperature optimum:
Fragment length <3 kb: Approximately +72°C
Fragment length >3 kb: Approximately +68°C
Substrates: Incorporates dNTP, dUPT, various labeled or modified nucleotides
Divalent ion requirement: Mg2+ (1.75 mmol/L when using 350 μmol/L of each dNTP; 2.75 mmol/L when using 500 μmol/L of each dNTP)
Recommended usage per 50 μL reaction: 0.5-5.0 U (3.75 U standard concentration)
Appearance: Clear, colorless solution
Storage buffer: Tris/HCl, 20 mmol/L; KCI, 100 mmol/L; EDTA, 0.1 mmol/L; DTT, 1 mmol/L; Nonidet P40, 0.5% (v/v); Tween 20, 0.5% (v/v); glycerol, 50% (v/v); pH approximately 8.0 at +4°C
Volume activity: ≥5 U/μL
Unspecific endonucleases (λDNA and MWM II DNA): Not detectable in up to 3 μL after 16 hours incubation at +65°C.
Nicking activity (pBR322 DNA): Not detectable in up to 3 μL after 16 hours incubation at +65°C.
Function test in PCR (human genomic DNA, 9.3,12, and 15 kb fragments, by using of 200 ng DNA positive): Corresponds to specification
Stability: At -15 to -25°C within specification range for 24 months.
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License disclaimer

48:Use of this product is covered by US patent claims and corresponding patent claims outside the US. The purchase of this product includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser’s own internal research. No right under any other patent claim, no right to perform any patented method, and no right to perform commercial services of any kind, including without limitation reporting the results of purchaser’s activities for a fee or other commercial consideration, is conveyed expressly, by implication, or by estoppel. This product is for research use only. Diagnostic uses under Roche patent claims require a separate license from Roche. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.