Ready-Lyse Lysozyme Solution is a stabilised lysozyme preparation for general bacterial lysis of Gram-positive or Gram-negative bacteria. Ready-Lyse’s specific activity is over 200 times higher than the commonly used egg-white lysozyme. This improves bacterial lysis and uses less enzyme, virtually eliminating losses due to nonspecific binding in nucleic acid purifications.
Complete digestion should occur within 15 minutes at room temperature. Following lysis, the lysate can be treated according to standard protocols for the purification of nucleic acids or proteins.
For difficult to lyse bacteria, Ready-Lyse Lysozyme Solution can be an additional lysis step when paired with our QuickExtract™ DNA Extraction Solution or MasterPure™ Gram Positive DNA Purification Kit, for DNA extraction. QuickExtract is formulated for bacterial DNA extractions, making it an optimal solution for DNA extraction from gram +/- bacteria, and our MasterPure Kit provides all of the reagents necessary to purify DNA from Gram-positive bacteria. Both QuickExtract and MasterPure can be used alone or with Ready-Lyse Lysozyme Solution.
The process is simple. Just add Ready-Lyse and QuickExtract to a bacteria sample and incubate at room temperature for 15 minutes. To kill remaining viable bacteria, the samples can be heated to 80 °C for 2 minutes.
Figure 1. Decreased loss of DNA with Ready-Lyse Lysozyme Solution compared to egg-white lysozyme.
pHC79 cosmid DNA (500 μg/mL) was incubated for 15 minutes at 22 °C with either 5 μg (30 KU)/mL of Ready-Lyse Lysozyme (RL), 500 μg/mL of egg-white lysozyme (EW) or no lysozyme (C) in conditions typically used for lysis of E. coli (25 mM Tris [pH 8.0], 10 mM EDTA). The solutions were then microcentrifuged for 10 minutes. The supernatants were removed and the pellets were resuspended in TE buffer containing 0.1% SDS. Supernatants (lanes 1-3) and pellets (lanes 5-7) were then analysed by electrophoresis in a 0.8% agarose gel.
The DNA extracted can be used for PCR, pulse-field gel electrophoresis (PFGE), restriction digestion, or optical mapping.
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Key features
- Highly Active: 200X more active than egg-white lysozyme and optimally active at a lower pH of 6.5-7.5 than the standard pH 9.
- Easy: No sample agitation or heat generation necessary during lysis which helps preserve protein function
- Flexible: Usage volumes are easily adjusted based on the scale of the experiment
- Stable: stable at -20 °C, eliminating the need to prepare a fresh solution for each use
- Accurate: virtually no nonspecific binding and significantly less precipitation of DNA compared to egg-white lysozyme
Specifications
Unit Definition: One unit of Ready-Lyse Lysozyme produces a decrease in A 350 of 0.001 per minute at 23 °C with a 0.5 mg/mL suspension of lyophilised E. coli K802 cells in 50 mM Tris-HCl (pH 7.5).
Storage Buffer: 50% glycerol containing 50 mM Tris-HCl (pH 7.5), 0.1 M NaCl, 0.1 mM EDTA, 1 mM DTT, and 0.1% Triton® X-100.
Product Details
Ready-Lyse Lysozyme Solution a ready-to-use solution supplied in quantities of 4 or 10 × 106 units.
Expert Recommendation: For difficult to lyse bacteria, Ready-Lyse Lysozyme Solution can be an additional lysis step when paired with our QuickExtract™ DNA Extraction Solution or MasterPure™ Gram Positive DNA Purification Kit, to help extract PCR-ready DNA from gram +/- bacteria or fungi. Both QuickExtract and MasterPure can be used alone or with Ready-Lyse Lysozyme Solution.
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