Zymo Research Zymoprep Yeast Plasmid Miniprep I

Zymo Research Zymoprep Yeast Plasmid Miniprep I Kit – Reagent Lysis and Isopropanol Precipitation Format, High-Efficiency Zymolyase Digestion, Bead-Free Isolation, Research Grade (MPN: D2001)

The Zymo Research D2001 (also cataloged as ZD2001 or ZD2001-EA) is a highly efficient molecular-biology-grade yeast plasmid purification kit engineered to rescue low-copy and hard-to-isolate plasmids directly from fungal cultures. Based on a modified alkaline lysis protocol, this kit utilizes a specialized formulation of Zymolyase yeast lytic enzyme as the primary digestion solution. This targeted enzymatic action breaks down the resilient glucan cell walls of yeasts without requiring cumbersome glass beads, mechanical vortexing, or hazardous phenol-chloroform extractions. Optimized for an alcohol precipitation format using isopropanol, the kit features a flexible elution timeline yielding pure plasmid DNA in standard TE buffer. This extracted DNA is fully prepared for immediate downstream applications, including high-efficiency E. coli transformation, polymerase chain reaction (PCR) amplification, Western blotting, and Southern hybridization assays.

Technical Performance and Method Protocols

• Enzymatic Zymolyase Cell Wall Digestion: Employs high-purity Zymolyase lytic enzymes to cleanly digest the polysaccharide matrix of yeast cell walls. This gentle enzymatic breakdown keeps the delicate internal plasmid strands physically intact, leading to significantly higher recovery rates compared to rough bead-beating methods.

• Bead-Free and Phenol-Free Lysis Pathway: Replaces mechanical glass-bead shaking and toxic organic solvents with a clean, room-temperature chemical and enzymatic alkaline lysis cascade. This workflow reduces counter friction, eliminates mechanical DNA shearing, and shields the technician from hazardous chemical exposures.

• Isopropanol Precipitation Isolation Format: Utilizes a classic, robust alcohol precipitation phase instead of spin columns. The cleared supernatant combines directly with isopropanol to precipitate the target nucleic acids, allowing technicians to collect the final pellet and resuspend it to their exact downstream concentration requirements.

• Multi-Format Sample Harvesting: Natively cleared to process up to 1.5 mL of liquid cell cultures, cell patches scraped directly from agar growth plates, or isolated single yeast colonies, giving researchers versatile harvesting options across variable library screen sizes.

Product Specifications

• Catalog Number / MPN: D2001 (Cross-listed as ZD2001 / D2001-EA)

• Manufacturer / Brand: Zymo Research Corporation

• Kit Method Classification: Zymoprep Yeast Plasmid Miniprep I (Isopropanol Precipitation / Reagent Format)

• Target Plasmid Size Window: Validated for stable DNA vector recovery up to 23 kb

• Expected Material Yields: Typically isolates 0.01 ng to 0.3 ng of DNA for standard 2 µm-based yeast plasmids from a standard 1.5 mL overnight culture format

• Downstream Minimum Elution Baseline: Recommends a minimum resuspension volume of 35 µL using the provided TE Buffer or molecular-grade water

• Total Process Execution Timeline: Under 30 minutes total operating duration (excluding baseline enzyme digestion incubation loops)

Key Benefits and Applications

• Target Organism Wall Vulnerability: The optimized lytic enzyme choice provides comprehensive wall digestion across Saccharomyces cerevisiae, Candida albicans, Schizosaccharomyces pombe, Pichia pastoris, and other fungi susceptible to Zymolyase cell wall processing.

• Low-Copy Plasmid Rescue: The highly sensitive chemical precipitation loop efficiently handles low-copy cloning vectors, helping researchers capture intact genetic sequences from tough industrial yeast strains and primary genomic screens.

• Downstream E. coli Transformation: Recovered plasmid eluate is free of chemical inhibitors, allowing it to be transformed directly into competent E. coli cells for rapid amplification, verification, and long-term library storage.

• Targeted Component Temperature Tuning: Kit includes distinct thermodynamic storage reagents, grouping ambient room-temperature buffers alongside a separate lyophilized Zymolyase vial optimized for sub-zero (-20 C) enzyme stability storage cascades.

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