| Product Name |
Thermostable MutS Mismatch Repair Protein (6×His-MBP-Taq-MutS Fusion)
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| Catalog Number |
[Your catalog number here] |
| Size |
100 µg (1 mg/mL stock concentration) |
| Description |
Recombinant Thermus aquaticus MutS protein expressed as an N-terminal 6×His-MBP fusion for enhanced solubility and purification. This thermostable mismatch-binding protein specifically recognizes and binds to various DNA mismatches with high affinity across a broad temperature range (4–70°C).
Key Features:
- Recognizes all single base mismatches (GT, CT, AG, etc.) and 1-4 bp indels
- Maintains activity from 0–75°C with optimal binding at 65°C
- Exhibits thermostable ATPase activity (kcat = 0.5 min-1 at 65°C)
- Dual-affinity tags (6×His and MBP) enable multiple purification strategies
- >95% purity by SDS-PAGE analysis
|
| Applications |
- Error correction in synthetic DNA and gene synthesis workflows
- High-throughput mutation detection and SNP screening
- Isothermal nucleic acid amplification quality control
- DNA heteroduplex analysis and mismatch characterization
|
| Source |
Recombinant expression in E. coli |
| Protein Characteristics |
- MW: 135.9 kDa (theoretical)
- Extinction Coefficient: 149,430 M-1cm-1 (280 nm)
- Fusion Tags: N-terminal 6×His and maltose-binding protein (MBP)
- Accession: AAC43637 (T. aquaticus MutS)
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| Quality Control |
- >95% purity by SDS-PAGE (Coomassie staining)
- Functional validation using standardized mismatch-binding assay
- Endotoxin level: <0.1 EU/µg
- Sterility tested (bacterial culture)
|
| Buffers |
Storage Buffer:
20 mM Tris-HCl (pH 8.0), 250 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 50% glycerol
Reaction Buffer (10×):
100 mM KCl, 50 mM Tris-HCl (pH 8.5), 10 mM MgCl2, 0.1 mM EDTA, 1 mM DTT, 2% glycerol
|
| Storage & Handling |
- Long-term: -80°C (stable for 24 months)
- Short-term: -20°C (stable for 6 months)
- Shipping: Dry ice
- Note: Avoid repeated freeze-thaw cycles
|
| Example Protocol |
For error correction in gene synthesis:
- Purify PCR products (QIAquick kit) and elute in 10 mM Tris-HCl (pH 7.8)
- Denature/reanneal (95°C for 5 min, cool at 0.1°C/s to 25°C)
- Add 1× reaction buffer and Taq MutS (950 nM final concentration)
- Incubate 10 min at room temperature
- Add amylose resin (NEB) and incubate 30 min with gentle mixing
- Collect supernatant for downstream processing
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| References |
- Carr PA, et al. (2004) Protein-mediated error correction for de novo DNA synthesis. Nucleic Acids Res 32(20):e162. doi:10.1093/nar/gnh160
- Matzas M, et al. (2005) Correcting errors in synthetic DNA through consensus shuffling. Nucleic Acids Res 33(6):e55. doi:10.1093/nar/gni054
- Bui CT, et al. (2007) Rapid SNP diagnostics using asymmetric isothermal amplification. Nat Methods 4:257-262. doi:10.1038/nmeth1007
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