T4 Gene 32 single-stranded DNA (ssDNA) binding protein

T4 Gene 32 Protein is an essential single-stranded DNA (ssDNA) binding protein that plays multiple roles in bacteriophage T4 DNA metabolism. Key characteristics include:

• Molecular Weight: 33.5 kDa (monomeric form)
• Binding Capacity: Binds cooperatively to ssDNA with a stoichiometry of 1 protein per 7-10 nucleotides
• Thermostability: Remains active up to 45°C
• Functional Domains: Contains distinct ssDNA binding and protein interaction domains
• Buffer Compatibility: Active in most common molecular biology buffers (pH 6.5-8.5)

• Nucleic Acid Manipulation:
  • Stabilization of ssDNA structures for electron microscopy
  • Prevention of ssDNA secondary structure formation
  • Enhancement of DNA hybridization efficiency
• Enzyme Reactions:
  • Increase yield and processivity in RT-PCR (0.1-1 μg/reaction)
  • Enhance T4 DNA polymerase activity (1:1 molar ratio with polymerase)
  • Improve PCR specificity and yield (0.5-2 μg/50 μL reaction)
  • Facilitate restriction enzyme digestion of problematic templates

T4 Gene 32 Protein: 10 mg/mL in storage buffer (10 mM Tris-HCl, 300 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 50% glycerol, pH 7.5 @ 25°C)

• Tested for absence of endonuclease, exonuclease, and phosphatase activities
• Verified ssDNA binding activity by electrophoretic mobility shift assay
• Confirmed >95% purity by SDS-PAGE analysis
• Functionally validated in PCR enhancement assays

• Short-term: -20°C (stable for 12 months)
• Long-term: -80°C (stable for 24 months)
• Note: Aliquot to avoid repeated freeze-thaw cycles

Typical working concentrations:

• PCR/RT-PCR: 0.1-1 μg per 50 μL reaction
• ssDNA stabilization: 1-2 μg per μg DNA
• Restriction enzyme enhancement: 0.5-1 μg per reaction