# Rapid and Equipment-Free Readout of CRISPR-Cas12/13 Assays Using Due Bio LF-detect Strips
## Abstract
Traditional CRISPR-Cas12/13 detection methods rely on complex laboratory equipment and time-consuming fluorescence reading systems, limiting their deployment in resource-constrained settings. Due Bio’s LF-detect lateral flow strips provide a simple, visual readout solution that delivers clear results within 15 minutes—no instruments required. As a leading POCT manufacturer, Due Bio enables researchers and diagnostic developers to transform CRISPR assays into field-deployable tests using standardized lateral flow strips compatible with Cas12/13 reporter systems.
## Introduction
CRISPR-Cas12 and Cas13 technologies have revolutionized molecular diagnostics by offering sequence-specific nucleic acid detection with exceptional sensitivity and specificity. However, the widespread adoption of CRISPR-based diagnostics in point-of-care (POC) settings has been hindered by the need for specialized equipment to detect fluorescence signals or interpret amplification curves.
Point-of-Care Testing demands rapid, equipment-free, and visually interpretable results—especially in low-resource environments, field epidemiology, and decentralized testing scenarios. Due Bio’s LF-detect Cas12/13 Dedicated Nucleic Acid Lateral Flow Strips address this critical gap by providing a simple chromatographic readout platform that integrates seamlessly with CRISPR cleavage reactions.
## Materials Required
– **Due Bio LF-detect: Cas12/13 Dedicated Nucleic Acid Lateral Flow Strips** (Cat. No. JY0301)
– **Due Bio IVD Reagents** (CRISPR-Cas12/13 reaction buffers and enzymes)
– Biotin-labeled and FAM-labeled reporter oligonucleotides
– Target nucleic acid samples (DNA or RNA)
– CRISPR-Cas12 or Cas13 enzyme
– Reaction tubes or 96-well microtiter plate
– Micropipettes and filtered pipette tips
– Timer or stopwatch
*Note: All Due Bio LF-detect strips are supplied ready-to-use (50 strips/tube) in moisture-proof aluminum foil packaging. Store at 4–30°C in a dark, dry place.*
## Step-by-Step Protocol
### 1. Sample Preparation
Extract nucleic acid from your sample using your preferred method (commercial kits, boiling lysis, or rapid extraction buffers). Ensure final DNA/RNA concentration is within the optimal range for CRISPR detection (typically 1–100 ng/μL). For RNA targets, include a reverse transcription step if using Cas12.
### 2. CRISPR Cleavage Reaction
Prepare the CRISPR reaction mix by combining:
– Cas12 or Cas13 enzyme
– crRNA/tracrRNA complex (target-specific)
– Biotin-FAM dual-labeled reporter oligonucleotide
– Reaction buffer (Due Bio IVD Reagents)
– Prepared sample (target nucleic acid)
Incubate at 37°C for 15–30 minutes to allow target-activated Cas enzyme cleavage of the reporter molecule.
### 3. Strip Incubation
Remove the required number of LF-detect strips from the tube. Immediately reseal the tube cap tightly to protect remaining strips from moisture. Add 50–100 μL of the cleavage reaction mixture directly to the sample well of the lateral flow strip. No additional buffer is required.
### 4. Result Interpretation
Wait 10–15 minutes for the strip to develop. Read results under normal lighting conditions:
– **Positive Result**: Control line (C) only — indicates Cas enzyme cleaved the reporter, preventing test line capture.
– **Negative Result**: Both Control (C) and Test (T) lines visible — intact reporter is captured at test line.
– **Invalid Result**: No control line — repeat the test with a new strip.
### 5. Documentation and Disposal
Photograph results for record-keeping within 30 minutes of development. Dispose of used strips according to local biohazard regulations. Do not interpret results after 30 minutes, as background signal may increase.
## Troubleshooting
### Q: I see faint or no control line. What could be wrong?
**A**: This typically indicates insufficient sample volume or excessive moisture exposure. Ensure you add at least 50 μL of reaction mixture to the sample well. Always store strips in their original moisture-proof packaging and reseal immediately after use. If the problem persists, dry unused strips at room temperature (18–28°C) for 8 hours before testing.
### Q: The test line appears very weak even for negative controls. Why?
**A**: Weak test lines may result from suboptimal reporter concentration or degraded reagents. Verify that your Biotin-FAM reporter is properly labeled and stored at –20°C. Use fresh Due Bio IVD Reagents and ensure the CRISPR reaction incubation time is sufficient (minimum 15 minutes). If issues continue, contact our technical support team for batch-specific guidance.
### Q: Can I use these strips with LAMP or RPA amplification products?
**A**: Yes. LF-detect Cas12/13 strips are compatible with any amplification method (PCR, LAMP, RPA/RAA) as long as the CRISPR cleavage reaction is performed before strip application. The strips detect the cleaved reporter, not the amplification product directly.
## Conclusion & OEM/ODM Call-to-Action
Due Bio’s LF-detect Cas12/13 Dedicated Nucleic Acid Lateral Flow Strips transform complex CRISPR assays into simple, field-deployable diagnostic tests. With 15-minute visual readout, no equipment requirements, and seamless integration with standard CRISPR workflows, researchers and diagnostic developers can accelerate their path from bench to bedside.
**Established in 1987**, Due Bio is a global POCT manufacturer committed to advancing accessible molecular diagnostics. Beyond our standard LF-detect product line, we provide comprehensive **IVD Reagents OEM/ODM services** for partners worldwide—including custom reporter design, private labeling, and large-scale manufacturing under ISO 13485 quality systems.
Whether you’re developing a CRISPR-based infectious disease assay, a cancer mutation detection panel, or a veterinary diagnostic test, Due Bio’s technical team can support your project from prototype to commercialization.
**Contact us today**: medtiger@foxmail.com
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*For research use only. Not for diagnostic procedures unless validated and approved according to local regulations.*