CHO Residual DNA Detection Kit (qPCR)
$480.00 - $1,440.00
$1,800.00
All products have special prices for bulk purchase, please contact for more details if required.
Cat. No.: CRDDK-50 (for 50T)
Cat. No.: CRDDK-200 (for 200T)
Description
CHO Residual DNA Detection Kit (qPCR), also known as CHO Genomic DNA Residue Probe qPCR Kit or CHO Host Cell Genomic DNA Residue Detection Kit, is a kit that uses probe-based quantitative PCR (qPCR) to rapidly, efficiently, and sensitively detect and quantify the residual genomic DNA from CHO host cells in various biological product samples. This product is contamination-resistant, containing optimized proportions of high-quality UDG enzyme and dUTP to effectively eliminate false positives or low CT values caused by product contamination during PCR amplification.
Chinese hamster ovary (CHO) cells are a commonly used cell line capable of expressing complex proteins with glycosylation modifications and other post-translational modifications. They are highly adaptable, suitable for large-scale culture and recombinant protein expression and purification. Compared to mammalian cells, they have fewer chromosomes. CHO cells are widely used in genetics, toxicity screening, nutrition, and gene expression studies, and are currently the most commonly used mammalian host cells for industrial production of recombinant protein therapeutic drugs. Even with strict purification processes, biological products produced using CHO cells may still contain trace amounts of residual CHO cell DNA fragments. These residual DNA fragments may pose biological hazards, and when their residual levels are too high, they may increase the risks of inflammation, carcinogenicity, and immune regulation disorders. Many countries and regions have strict requirements and restrictions on residual host cell DNA in biopharmaceutical products. By detecting and controlling residual host cell DNA, the immunogenicity risk and safety of biological products can be assessed.
Components
- Probe qPCR Mix (2X, UDG)
- Primer/Probe Mix (10X)
- Ultrapure Water
- Dilution Buffer
- CHO DNA Control (30 ng/μl)
- Low ROX (50X)
- High ROX (50X)
Features
- This kit can efficiently and sensitively detect residual DNA from CHO host cells, with a minimum detection limit as low as femtograms (fg). Based on the quantitative PCR method for exogenous DNA residue measurement, this kit uses probe-based qPCR with a detection limit of 3fg/μl and a detection range of 3fg/μl to 300pg/μl.
- This kit is highly specific; HeLa cell and E. coli cell genomic DNA do not interfere with CHO DNA residue detection. The kit only detects CHO host cell DNA and shows no cross-reactivity with DNA from other species.
- This kit is highly stable, with fragmented DNA of varying degrees having minimal interference on the detection of CHO DNA residues. By using different amounts of Micrococcal Nuclease (MNase) to fragment the CHO DNA Control to varying extents, and then performing qPCR detection with this kit, it was shown that DNA fragments longer than 200bp are mostly detected without affecting the detection results. However, if the fragmentation is too severe, resulting in a predominance of DNA fragments less than 200bp, the detected residual DNA amount may be lower.
- This kit has good repeatability. The CT values for 12 repeated tests detecting 3pg/μl and 30fg/μl CHO DNA are shown in the table below, with CV values (coefficient of variation) all being less than 5%.
- This kit can be used with the Residual DNA Sample Preparation Kit with Magnetic Beads for sample extraction. The recovery test results using the CHO DNA Control as a simulated sample are shown in the table below.
Note 1: Recovery rates may vary depending on sample concentration, volume, sample type, and experimental procedures. The data in the table is for reference only.
Note 2: According to the "Determination Method for Exogenous DNA Residue" in the Chinese Pharmacopoeia (2020 Edition, Volume III) "Third Method: Quantitative PCR," the recovery rate of spiked samples should be between 50% and 150%.
This kit provides Probe qPCR Mix (2X, UDG) and specific primer-probe mix required for qPCR experiments. The premix includes high-quality, CHO DNA-free Taq DNA Polymerase and UDG enzyme, PCR Buffer, dNTPs, dUTP, stabilizers, and magnesium ions—all general components. The high-purity primer-probe mix targets highly specific sequences in the CHO genome, allowing for ultra-high sensitivity quantitative detection of CHO DNA. Additionally, this kit includes CHO DNA Control, serving as a positive control to verify the kit’s functionality and to construct standard curves.
Storage
Store at -20ºC for one year. Primer/Probe Mix (10X), Low ROX (50X), and High ROX (50X) should be stored protected from light and avoid repeated freeze-thaw cycles.
Precautions
- It is recommended to use this product immediately after first opening, or to properly aliquot and store it at -20ºC under sterile conditions to prevent contamination by airborne E.coli, which could affect experimental results.
- Ensure that the reagents are completely thawed and thoroughly mixed before use. Avoid generating bubbles during mixing.
- Primer/Probe Mix (10X), Low ROX (50X), and High ROX (50X) contain fluorescent dyes. Avoid strong light exposure when storing this product or setting up the PCR reaction system to prevent fluorescence quenching.
- Tests have shown that repeated freeze-thaw cycles up to 10 times do not significantly affect the product’s performance, but it is still recommended to avoid repeated freeze-thaw cycles as they may degrade product performance.
- qPCR detection is highly sensitive; it is recommended to perform the detection in a standard PCR laboratory. The PCR reaction setup area should be free from potential amplification product contamination. Although this product is contamination-resistant, it is still advised not to tear open the PCR sealing film or open the PCR tube cap in the PCR reaction setup area. PCR products should be sealed and handled according to post-amplification product requirements to avoid contamination of the experimental environment with high concentrations of PCR products.
- It is recommended to use filter tips to prepare the PCR system to minimize false positives caused by contamination.
- It is recommended to use low-adsorption 1.5 ml centrifuge tubes for high sample recovery rates.
- This product is for scientific research use only by professionals and is not intended for clinical diagnosis or treatment, food, or drugs, and should not be stored in ordinary residential areas.
- For your safety and health, please wear a lab coat and disposable gloves during operation.
Related:
- E.coli Residual DNA Detection Kit (qPCR)
- CHO Residual DNA Detection Kit (qPCR)
- Residual DNA Sample Preparation Kit with Magnetic Beads
Only for research and not intended for treatment of humans or animals
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