HCY™ 96 DNA RNA Extration Kit

Of course. Let me provide you with detailed information about the HCY™ 96 DNA/RNA Extraction Kit.

This is a high-throughput nucleic acid extraction kit designed for use in laboratories that process a large number of samples, such as in clinical diagnostics, pathogen detection, and genomic research.

Here is a comprehensive breakdown of its features, principle, typical procedure, and applications.

Overview

The HCY™ 96 DNA/RNA Extraction Kit is a magnetic bead-based purification system formatted for 96-well plates. It allows for the simultaneous purification of both DNA and RNA from a single sample in a automated or semi-automated workflow. This is crucial for applications like pathogen detection (e.g., SARS-CoV-2, influenza, respiratory syncytial virus), where testing for multiple targets from one sample is common.

Key Features

1. High-Throughput: Designed for 96-well plates, making it ideal for processing 96 samples simultaneously. It is compatible with automated liquid handling platforms and magnetic bead processors.
2. Dual Extraction: Efficiently co-extracts both genomic DNA and total RNA from a single sample lysate. This saves time, reagents, and sample material.
3. Magnetic Bead Technology: Uses a silica-based magnetic bead matrix to bind nucleic acids. This method is known for its high purity, yield, and ease of automation compared to traditional spin-column methods.
4. High Purity and Yield: The optimized buffers and wash steps remove inhibitors like proteins, salts, and other contaminants, resulting in high-quality DNA and RNA suitable for sensitive downstream applications.
5. Rapid Procedure: The entire process can be completed in 20-40 minutes for a full 96-well plate, depending on the automation equipment used.
6. Wide Sample Compatibility: Can be used with a variety of sample types, including:
   • Swab samples (nasopharyngeal, oropharyngeal)
   • Serum / Plasma
   • Cell culture supernatants
   • Tissue homogenates
   • Bodily fluids

Principle of the Method

The kit operates on the principle of silica-magnetic bead nucleic acid purification:

1. Lysis: The sample is lysed with a strong chaotropic salt buffer (usually containing Guanine-HCl or Guanine Thiocyanate). This buffer denatures proteins, inactivates nucleases (RNases and DNases), and releases nucleic acids (DNA and RNA) into solution.
2. Binding: In the presence of the lysis buffer and a binding solution, nucleic acids specifically bind to the surface of the silica-coated magnetic beads. The chaotropic salts disrupt the water structure, allowing the negatively charged phosphate backbone of the nucleic acids to interact with the positively charged silica surface.
3. Capture & Washes: A magnetic stand (or a magnetic rod in an automated system) is used to immobilize the bead-nucleic acid complexes against the wall of the tube/plate. The supernatant is removed, and the beads are washed 2-3 times with ethanol-containing wash buffers. These washes remove salts, proteins, lipids, and other impurities without eluting the nucleic acids.
4. Elution: After a final wash and complete drying to remove residual ethanol, the pure nucleic acids are eluted from the beads using a low-salt buffer (e.g., TE Buffer or Nuclease-Free Water). The elution buffer disrupts the interaction between the silica and the nucleic acids, releasing them into the final clean solution.

Typical Workflow (Simplified)

1. Prepare Lysate: Mix the sample with Proteinase K (if required) and Lysis Buffer. Incubate to complete lysis.
2. Bind: Transfer the lysate to a 96-well deep-well plate. Add magnetic beads and Binding Solution. Mix thoroughly to allow nucleic acids to bind to the beads.
3. Capture: Place the plate on a magnetic stand until the solution clears. Aspirate and discard the supernatant.
4. Wash 1: Add Wash Buffer 1, mix to re-suspend the beads, capture on the magnet, and discard the supernatant.
5. Wash 2: Add Wash Buffer 2, mix, capture, and discard the supernatant.
6. Dry: Air-dry the bead pellet for a few minutes to ensure all ethanol from the wash buffer has evaporated.
7. Elute: Add Elution Buffer, mix thoroughly, incubate for a few minutes, and capture the beads. The supernatant now contains the purified DNA/RNA and is transferred to a new plate or tube.

Downstream Applications

The high-quality DNA and RNA extracted with this kit are suitable for a wide range of sensitive molecular biology applications:

• Real-Time RT-PCR (qRT-PCR): The primary application, especially for viral detection (e.g., COVID-19 testing).
• PCR
• Next-Generation Sequencing (NGS)
• Microarray Analysis
• Cloning
• Molecular Diagnostics

Summary

The HCY™ 96 DNA/RNA Extraction Kit is a powerful tool for modern molecular laboratories that require efficiency, scalability, and reliability. Its magnetic bead, 96-well format is the industry standard for high-throughput diagnostic and research labs, enabling fast and accurate preparation of nucleic acids for critical downstream analyses.

Disclaimer: For the most accurate and specific protocol, always refer to the official manufacturer’s instructions provided with the kit.