SEQQX® OnePot Pro DNA Library Prep Kit
An enzymatic DNA library preparation kit for Illumina and MGI sequencing, combining fragmentation, end repair and dA-tailing in one tube for 1 ng–1 µg DNA input.
One-tube fragmentation module
Illumina and MGI
RUO
Product overview
SEQQX® OnePot Pro DNA Library Prep Kit, catalog no. LM801003, prepares sequencing libraries from genomic DNA, full-length cDNA and similar samples. Enzymatic fragmentation replaces sonication, while fragmentation, end repair and dA-tailing are combined in one tube. The kit includes ligation reagents and a high-fidelity amplification mix and works with platform-matched adapters and primers for Illumina® or MGI® sequencing.
Integrated fragmentation module
Combine DNA fragmentation, end repair and dA-tailing in one tube without a separate sonication step.
Flexible DNA input
The handbook specifies 1 ng–1 µg input and supports animal, plant and microbial genomic DNA as well as full-length cDNA.
Platform and format options
Use matching Illumina® or MGI® adapters and primers; select standard packs or a 96 T automation-ready plate format.
Specifications
| Parameter | Value |
|---|---|
| Method | Enzymatic DNA fragmentation and sequencing library preparation |
| Input amount | 1 ng–1 µg DNA |
| Sample types | Animal, plant or microbial genomic DNA; full-length cDNA |
| Platforms | Illumina® and MGI® with matching adapters and primers |
| Integrated module | Fragmentation, end repair and dA-tailing in one tube |
| Fragmentation reaction | 60 µL; 10 µL each of Smearase® Buffer 4.0 and Enzyme 4.0 |
| Standard incubation | 4°C for 1 min; 30°C for 10–40 min; 72°C for 20 min |
| Adapter ligation | 110 µL reaction; 20°C for 15 min; heated lid off |
| Amplification | 50 µL reaction with 2× Ultima HF Amplification Mix |
| Additional reagents | Platform-matched adapters/primers and DNA clean-up beads |
| Library QC | Fluorescence or qPCR quantification; capillary electrophoresis for size |
| Storage and shelf life | −25°C to −15°C; up to 12 months |
| Intended use | Research use only; not for diagnostic or clinical procedures |
Pack sizes & ordering
| Catalog No. | Pack size | Format |
|---|---|---|
| LM801003ES01 | 8 T | Standard kit |
| LM801003ES02 | 24 T | Standard kit |
| LM801003ES03 | 96 T | Standard kit |
| LM801003ES04 | 96 T | Automation-ready plate format |
One test (T) corresponds to one library-preparation reaction; specify the required pack size and standard or automation-ready plate format when ordering.
Performance & validation
Bioanalyzer analysis of 250 ng standard genomic DNA shows that adjusting fragmentation time and temperature shifts the insert-size distribution under the tested conditions.
Time-dependent fragmentation
At 30°C, reference insert sizes shift from approximately 700 bp at 10 min to 150–200 bp at 40 min.
Temperature-dependent control
At 35°C, reference insert sizes shift from approximately 500–600 bp at 10 min to 130 bp at 40 min.
Electropherogram evidence
Figures 2 and 3 show Bioanalyzer traces after 2.0× bead purification. Optimize time and temperature for the sample and desired insert size.
Product documents
Product FAQs
Which DNA inputs are supported?
The handbook specifies 1 ng–1 µg of genomic DNA, full-length cDNA or similar samples, including animal, plant and microbial genomic DNA.
Does OnePot mean the entire workflow is a single step?
No. Fragmentation, end repair and dA-tailing share one tube. Adapter ligation, clean-up, amplification and final QC follow.
Are adapters and all primers included?
Adapters are not listed as kit components. Short adapters require the index primers from the matching adapter kit; long adapters require a separately ordered, platform-matched Primer Mix.
How is insert size adjusted?
Adjust fragmentation time and temperature. For high-quality genomic DNA, 30°C for 15 min is a starting point for approximately 300–500 bp inserts; optimize for your sample.
How should the kit be stored?
Store all components at −25°C to −15°C for up to 12 months. Research use only.