Unit Definition: One unit is defined as the amount 3. Mg++ and additives: 25 µl 50 µl FInal ® ++ LongAmp Taq of enzyme that will incorporate 10 nmol of dNTP COMPONENT REacTION REacTION CONCENTRATION Mg concentration of 1.5–2.0 mM is optimal into acid insoluble material in 30 minutes at 75°C. 5X LongAmp Taq for most PCR products generated with ++ DNA Polymerase Reaction Buffer 5 µl 10 µl 1X LongAmp Taq DNA Polymerase. The final Mg ™ Unit Assay Conditions: 1X ThermoPol Reaction 10 mM dNTPs 0.75 µl 1.5 µl 300 µM concentration in 1X LongAmp Taq Reaction Buffer, 200 µM dNTPs including [3H]-dTTP and Buffer is 2 mM. This supports satisfactory 1-800-632-7799 10 µM Forward Primer 1 µl 2 µl 0.4 µM (0.05–1 µM)
[email protected] 200 µg/ml activated Calf Thymus DNA. amplification of most amplicons. However, www.neb.com 10 µM Reverse Primer 1 µl 2 µl 0.4 µM (0.05–1 µM) Mg++ can be further optimized in 0.5 or 1.0 M0323S 010121214121 Heat Inactivation: No LongAmp Taq 5 units/ mM increments using MgSO . DNA Polymerase 1 µl 2 µl 50 µl PCR 4 Quality Control Assays Amplification of some difficult targets, like M0323S Template DNA variable variable <1,000 ng GC-rich sequences, may be improved Long Amplicon PCR: LongAmp Taq DNA Poly- Nuclease-Free Water to 25 µl to 50 µl 500 units 2,500 U/ml Lot: 0101212 merase is tested for the ability to amplify a 30 kb with additives, such as DMSO (4) or amplicon from lambda DNA and a 30 kb amplicon Notes: Gently mix the reaction.