Expand Long Template PCR System
For customers in the European Economic Area: Contains SVHC: octyl/nonylphenol ethoxylates. For further processing on its own or in a mixture as part of an IVD method and under controlled conditions only – acc. to Art. 56 (3) and 3 no. 23 REACH Regulation.
collapse
| -
Expand Long Template PCR System material number and pack size: Material Number Pack Size 03321053103 custom fill
The enzyme is supplied without reaction buffer. -
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.
- Amplify long templates.
- 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
-
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.