Biofilm DNA Extraction Kit, IC-5339
Product Introduction
This product is a specialized spin-column based kit designed for the rapid and efficient isolation of high-quality genomic DNA from biofilm samples. Biofilms present unique challenges for DNA extraction due to the complex extracellular polymeric substance (EPS) matrix composed of polysaccharides, proteins, and extracellular DNA that encapsulates the microbial community. The kit employs an optimized lysis buffer system combined with mechanical disruption to effectively penetrate and dissolve the biofilm matrix, releasing DNA from embedded bacteria, fungi, and other microorganisms.
The extracted DNA is purified through a silica-based spin column that selectively binds nucleic acids while removing proteins, polysaccharides, humic acids, and other PCR inhibitors commonly found in environmental and clinical biofilm samples. The purified DNA is suitable for downstream applications including quantitative PCR, next-generation sequencing, metagenomic analysis, and 16S rRNA gene amplification.
Product Features
1. Specialized for Biofilm: Optimized lysis chemistry effectively disrupts the recalcitrant EPS matrix of biofilms.
2. Efficient Inhibitor Removal: Proprietary buffer system removes polysaccharides, humic substances, and other PCR inhibitors to ensure high-quality DNA suitable for sensitive enzymatic reactions.
3. High DNA Yield and Purity: Silica membrane technology delivers pure DNA with A260/A280 ratios between 1.7 and 1.9.
4. Fast and Convenient: Complete extraction protocol can be performed in under 60 minutes.
5. Versatile Input: Compatible with biofilms collected from medical devices, industrial pipelines, natural aquatic surfaces, and laboratory flow cells.
Specifications
Size: 50 Preps
Sample Input: Up to 200 mg of biofilm biomass per preparation
Storage and Stability
Storage Conditions: Store kit components at room temperature (15-25¡æ). After reconstitution of Proteinase K, store at -20¡æ.
Shelf Life: The product is stable for 12 months from the date of manufacture when stored as directed.
Protocol (For Reference Only)
Important: Ensure all buffer concentrates have been reconstituted with ethanol or isopropanol according to the bottle labels before first use.
1. Sample Collection and Preparation: Collect biofilm sample by scraping from surfaces or centrifuging liquid suspensions, transfer up to 200 mg of biofilm biomass to a 2 mL microcentrifuge tube containing lysis buffer and sterile glass beads.
2. Mechanical Lysis: Vortex the tube vigorously at maximum speed for 10-15 minutes to physically disrupt the biofilm matrix, alternatively use a bead beater for more consistent homogenization.
3. Enzymatic Digestion: Add Proteinase K solution to the lysate, mix thoroughly, and incubate at 56¡æ for 15-30 minutes with periodic vortexing until the sample is completely solubilized.
4. Binding: Add binding buffer and ethanol to the lysate, mix by inversion, transfer the mixture to the spin column, and centrifuge at 10,000 x g for 1 minute, discarding the flow-through.
5. Washing: Wash the spin column membrane sequentially with Wash Buffer I and Wash Buffer II by centrifugation at 10,000 x g for 1 minute each, discarding flow-through after each wash.
6. Elution: Place the spin column in a clean 1.5 mL microcentrifuge tube, add 50-100 µL of pre-warmed (60¡æ) Elution Buffer directly to the membrane center, incubate at room temperature for 2 minutes, and centrifuge at 10,000 x g for 1 minute to collect purified DNA.
Precautions
1. Biofilm samples should be processed immediately after collection or stored at -80¡æ to prevent microbial community shifts and DNA degradation.
2. Complete mechanical disruption of the biofilm matrix is critical for optimal DNA yield; vortexing with beads should result in a homogeneous suspension without visible clumps.
3. Extended Proteinase K digestion may be necessary for particularly recalcitrant or thick biofilm samples.
4. Pre-warming the Elution Buffer to 60¡æ significantly improves DNA recovery from the silica membrane.
5. For research use only. Not for use in diagnostic or therapeutic procedures.
FAQ (Simplified)
Q1: My DNA yield is low. What could be the problem?
A1: Insufficient mechanical disruption is the most common cause. Ensure vigorous vortexing with beads or use a bead beater. Alternatively, the biofilm sample may be too large, exceeding the binding capacity of the column.
Q2: The extracted DNA appears brown or discolored. Is it still usable?
A2: Discoloration indicates the presence of co-purified humic substances or other inhibitors. Repeat the extraction with fresh sample and include an additional wash step with Wash Buffer II. Consider reducing the sample input amount for inhibitor-rich biofilms.
Q3: Can this kit be used for water or planktonic bacterial samples?
A3: Yes, the kit is compatible with planktonic bacterial pellets and water filter concentrates. For pure liquid cultures, a standard bacterial genomic DNA kit may be sufficient, but this kit provides additional inhibitor removal capacity for challenging environmental samples.
Q4: Is the extracted DNA suitable for long-read sequencing?
A4: Yes. The gentle spin-column method produces high molecular weight DNA suitable for Oxford Nanopore and PacBio long-read sequencing platforms. Avoid excessive vortexing during the binding step to minimize DNA shearing.
Disclaimer
1. For Research Use Only. Not for use in diagnostic or therapeutic procedures.
2. Due to the variable nature of biological research, optimization of conditions using a small-scale pilot experiment is strongly recommended before processing valuable samples.
3. This warranty is limited to the replacement of the product. The manufacturer assumes no liability for incidental or consequential damages, including loss of samples or data.
4. Wear appropriate protective clothing and gloves when handling this product.
Ordering Information
Catalog Number: IC-5339
Product Name: Biofilm DNA Extraction Kit
Size: 50 Preps
Price: CNY ¥5600.00 / USD $560.00 / EUR €672.00 / JPY ¥100800.00
2023 Version
