Accuris Brand - The Fast Extraction PCR Kit features an exclusive single-tube solution that extracts genomic DNA in just 8 minutes. This is swiftly followed by PCR amplification utilizing our Hot Start Red Dye Master Mix, designed for superior endpoint PCR performance. Hot Start Red Dye Master Mix is a 2X formula that incorporates a non-reactive red dye, enabling direct gelloading. This kit offers versatility to suit different workflows: the PCR prepared genomic DNA can be directly used in endpoint PCR, or can be applied in real-time PCR using either SYBR Green or TaqMan Probe chemistries.
Each 100 reaction kit includes:- 1 tube of High Fidelity Hot Start Red Dye Master Mix (1.25mL)
- 1 tube of Fast Extraction Lysis Solution (1.25mL)
Applications: Extraction & Amplification of Tissue, Plant, Saliva, and Bacteria
The Accuris Brand Fast Extraction PCR Kit offers exceptional versatility and can be used to extract and amplify DNA from a wide range of sample types
• Compatible with animal tissue, plant, saliva, & bacterial samples
• Extraction & Amplification in less than 60 minutes
• Includes High Fidelity Hot Start Red Dye Master Mix to prevent the risk of nonspecific amplification & to visualize bands easily
• Process extracted sample via PCR with included mastermix
• Alternatively amplify via qPCR (qPCR mastermix not included)
Accuris™ Fast Extraction PCR Kit Description
The Accuris Fast Extraction PCR is engineered for the rapid, cost-efficient, and effective extraction and amplification of DNA from a wide array of solid tissues, such as mouse tails, ear punches, plant leaves, bacteria, and saliva. This kit is composed of a unique single-tube extraction solution that enables the release of genomic DNA in just 8 minutes. Following extraction, the PCR amplification process utilizes the Accuris High Fidelity Hot Start Master Mix, which is optimized for high-quality end-point PCR results. The inclusion of an inert red dye in the High Fidelity Hot Start Master Mix facilitates the direct loading of the PCR product onto a gel. For enhanced versatility in laboratory applications, the genomic DNA prepared with this kit is immediately ready for use in end-point PCR and real-time PCR assays using SYBR Green or TaqMan Probe chemistries.
Upon receipt, immediately store at -20°C. Avoid excessive freeze/thaw cycles. When stored as directed, this product will retain its activity for 12 months from date of receipt. The product may also be stored at 4°C for up to one month.
Limitations of Use
For research purposes only. Not intended\ for therapeutic or diagnostic use.
Quality Control
Accuris enzymes and reagents are tested under general assay conditions for activity, reproducibility, efficiency, heat activation, sensitivity, and absence of nuclease contamination and nuclease activity. This product is manufactured under a comprehensive quality management system, following ISO 9001:2008 standards.
General Guidelines
Template
➢ Tissue: Use 0.5 – 10 mg / 100uL Lysis Solution
➢ Plant: Use 2.0 – 10 mg / 100uL Lysis Solution
➢ Saliva: Use 10 – 20 uL / 100uL Lysis Solution
➢ E. Coli : Use 1 Colony (0.5 – 2mm) / 100uL Lysis Solution
Reaction conditions
Most reactions can be carried out at a temperature of 42°C for 30 minutes. When working with templates that have a high GC content (above 65%), the temperature can be increased, up to 55°C, to alleviate any problems associated with secondary structure.
Analysis
Results can be analyzed using agarose gel electrophoresis. The High Fidelity Hot Start Master Mix contains an inert red tracking dye which migrates at the rate of 600 bp and 350 bp DNA in 1% and 2% TAE agarose gels, respectively.
Accuris is not responsible for consequential or incidental damages, direct or indirect, resulting from use of this product. Accuris guarantees the performance of this product as described when used in accordance with these instructions.
Reaction Setup
Thaw Lysis Solution. Note: The Lysis Solution has a cloudy appearance.
Component 25 μL reaction Final concentration
2X HF Red Dye Mix 12.5 μL 1X
Forward Primer (10μM) 1.0 μL 400 nm
Reverse Primer (10μM) 1.0 μL 400 nm
DNA Extract 2.0 – 4.0uL
PCR grade water to 25uL final reaction volume
Protocol
1. The DNA extraction process can be carried out in 0.2 mL PCR Tubes or 0.5 mL/1.5
mL microcentrifuge tubes in a thermal cycler or heating block.
2. Thaw Lysis Solution. Please note that the Lysis Solution has a cloudy appearance.
3. Add your sample to a tube containing 100 μl of the Lysis Solution.
4. Vortex the tube containing the sample and the lysis solution for 15 seconds.
5. Transfer the tube to a heat block or a thermal cycler and prepare a DNA extract of
PCR-ready DNA as follows:
Cycles Temperature & Time Notes
1 65°C, 6 minutes Lysis
1 98°C, 2 minutes Protease Deactivation
1 4°C Cool on ice