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FAQs for PreCR™ Repair Mix Protocols for PreCR™ Repair Mix FAQs for Polymerases Technical Reference for Polymerases Enzyme Finder NEBcutter NEBuffer Chart Double Digest Finder Isoschizomers DNA Sequences and Maps REBASE β-Nicotinamide adenine dinucleotide (NAD+) ThermoPol Reaction Buffer Taq 2X Master Mix Taq DNA Ligase Taq DNA Polymerase with ThermoPol Buffer 2-Log DNA Ladder (0.1–10.0 kb) Deoxynucleotide Solution Mix Endonuclease IV Endonuclease VIII Fpg Quick-Load® 2-Log DNA Ladder(0.1-10.0 kb) T4 PDG (T4 Endonuclease V) Uracil-DNA Glycosylase (UDG)

Home > Products > Polymerases > PreCR > PreCR™ Repair Mix PreCR™ Repair Mix Catalog # Size Concentration Price Qty   M0309L 150 reactions   $520.00 M0309S 30 reactions   $130.00 Prices are in US dollars and valid only for US orders. Download: Technical Bulletin|MSDS PDF Suitable for PCR reactions, microarrays or other DNA technologies Easy-to-use protocols Does not harm template Description: The PreCR™ Repair Mix is an enzyme cocktail formulated to repair damaged template DNA prior to its use in the polymerase chain reaction (PCR), microarrays or other DNA technologies. PreCR is active on a broad range of DNA damages, including those that block PCR (e.g. apurinic/apyrimidinic sites, thymine dimers, nicks and gaps) and those that are mutagenic (e.g. deaminated cytosine and 8-oxo-guanine). In addition, it will remove a variety of moieties from the 3´end of DNA leaving a hydroxyl group. The PreCR Repair Mix will not repair all damages that inhibit/interfere with PCR. For example, it will not repair 8-oxo-7,8-dihydro-2'deoxyadenosines or fragmented DNA. In fact, the ligase present in the mix is very active at sealing nicks in DNA but does not ligate blunt ends or nicks near a mismatch effectively. This limits the possibility of chimeric gene formation as a product of ligation. The PreCR Repair Mix can be used in conjunction with any thermophilic polymerase. Table 1: Types of DNA Damage Figure 1: Repair of different types of DNA damage with the PreCR Repair Mix. The gel shows trial amplifications from damaged DNA that was either not treated (-) or treated (+) with the PreCR Repair Mix. Type of DNA damage is shown. Note: heat treated DNA is incubated at 99°C for 3 minutes. Marker M is the 2-Log DNA Ladder (NEB #N3200). Source: Each of the recombinant proteins present in the PreCR Repair Mix has been expressed in E. coli. Applications: Repair of DNA prior to its use as a template in PCR or other DNA technologies. Reagents Supplied: β-Nicotinamide adenine dinucleotide (NAD+) (100X) L1 Primer Mix (20 μM) PreCR Repair Mix * (1X) Purified BSA (10X) UV damaged Lambda DNA (0.5 ng/μl) ThermoPol Reaction Buffer (10X) Properties & Usage Heat Inactivation: No Reaction & Storage Conditions Reaction Conditions: 1X ThermoPol Reaction Buffer Incubate at 37°C. 1X ThermoPol Reaction Buffer: 20 mM Tris-HCl 10 mM (NH4)2SO4 10 mM KCl 2 mM MgSO4 0.1 % Triton X-100 pH 8.8 @ 25°C Storage Temperature: -20°C Notes Usage notes: * PreCR Repair Mix components: Taq DNA Ligase, Endonuclease IV, Bst DNA Polymerase, Fpg, Uracil-DNA Glycosylase (UDG), T4 PDG (T4 Endonuclease V) and Endonuclease VIII. Reaction Conditions: DNA template, 100 µM dNTPs, 1X ThermoPol Reaction Buffer, 1X NAD+ and 1 µl PreCR Repair Mix in a total reaction volume of 50 µl. Incubate at 37°C. 1X NAD+ Solution: 0.5 mM NAD+. dNTPs are not included with the PreCR Repair Mix. FAQs Will PreCR ligate my DNA fragments? How much DNA will PreCR repair? What are the liquid volumes of the PreCR Repair Mix Components? Will treating my DNA with the PreCR Repair Mix hurt my reaction? What is the sequence of the L1 primer mix? How is the damaged DNA that you test the PreCR Repair Mix against created? Can I buy this damaged DNA separately? Why does not my control reaction give a detectable amplicon? Can I buy any of the PreCR Repair Mix components separately? Will the PreCR Repair Mix blunt the ends of the DNA? Why don’t I see the expected band from my repaired template? Is my buffer compatible and which reaction should I use? Quality Control for Current Lot Quality control values for a specific lot can be found on the datacard which accompanies each product. Quality Assurance Statement: The PreCR repair mix was tested for the ability to rescue successful amplification from UV damaged Lambda DNA (NEB #N3011) and from an oxidized plasmid. The mix was also tested for the ability to cleave oligonucleotide substrates containing thymine glycol, deoxyuracil, or 8-oxo-guanine in the absence of dNTPs. The absence of dNTPs prevents full repair and allows the presence of specific repair-targeting activities to be evaluated. Reagents Sold Separately β-Nicotinamide adenine dinucleotide (NAD+) ThermoPol Reaction Buffer Companion Products Taq 2X Master Mix Taq DNA Ligase Taq DNA Polymerase with ThermoPol Buffer 2-Log DNA Ladder (0.1–10.0 kb) Deoxynucleotide Solution Mix Endonuclease IV Endonuclease VIII Fpg Quick-Load® 2-Log DNA Ladder(0.1-10.0 kb) T4 PDG (T4 Endonuclease V) Uracil-DNA Glycosylase (UDG)