QCdetect(TM) Residual DNA Detection Kit for CHO cells
- for Genetic Research
- Manufacturer :
- FUJIFILM Wako Pure Chemical Corporation
- Storage Condition :
- Keep at -20 degrees C.
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Comparison
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100Tests
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In stock in Japan |
Document
Kit component
Kit Contents
| 1 x PCR Master Mix | 2 x 1 mL |
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| DNA Dilution Buffer (DDB) | 1 x 10 mL |
| CHO Control DNA | 1 x 40 μL |
Overview
This product is a kit for quantifying genomic DNA from CHO cells by qPCR (probe method). It can be used for residual DNA testing for antibody pharmaceuticals and their manufacturing processes.
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Features
- Limit of detection: ≥ 0.0003 pg/test
- Limit of quantitation: ≥ 0.003 pg/test
- Calibration curve with high linearity
- Minimal inter-assay variability and high reproducibility
- Easy operation using the pre-mix buffer
- Less susceptible to contaminants in samples
- Contains an internal control
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Detection wavelength
CHO gDNA 520nm (e.g., FAM) Internal Control 554nm (e.g., HEX) Note) Internal Control is non-natural synthetic DNA.
Application
Calibration curve
qPCR of 0.0003 pg to 30,000 pg of CHO gDNA was performed using this kit to prepare a calibration curve (n = 3).
A calibration curve with very high linearity was obtained over a wide concentration range from 0.0003 pg to 30,000 pg.
| Pg/well | 0.0003 | 0.003 | 0.03 | 0.3 | 3 | 30 | 300 | 3000 | 30000 |
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| Ct | 37.639 | 34.822 | 31.546 | 28.117 | 24.809 | 21.337 | 18.079 | 14.726 | 11.506 |
| 37.107 | 34.981 | 31.506 | 28.324 | 24.906 | 21.416 | 18.084 | 14.744 | 11.467 | |
| 37.642 | 34.689 | 31.595 | 28.267 | 24.987 | 21.533 | 18.225 | 14.807 | 11.269 | |
| Average Ct | 37.462 | 34.830 | 31.549 | 28.236 | 24.901 | 21.428 | 18.129 | 14.759 | 11.414 |
| S. E | 0.30806 | 0.14652 | 0.04455 | 0.10713 | 0.08904 | 0.09853 | 0.0831 | 0.0424 | 0.12685 |
Detection of fragmented DNA
CHO gDNA was fragmented by sonication and detected by this kit to examine whether the fragmented CHO gDNA could be detected.
Fragmented gDNA could be detected with the same sensitivity as intact gDNA. A decrease in detection sensitivity was not observed, even for low concentrations of gDNA.
Example of use for a high-protein sample -combined with DNA Extractor® Kit-
CHO gDNA was spiked to a sample containing high concentration of γ-globulin, and DNA was extracted by DNA Extractor® Kit.
The obtained DNA was quantified with QCdetect™ Residual DNA Detection Kit for CHO cells, and the spike recovery rate was obtained.
CHO gDNA could be recovered with a high recovery rate, even when a high-protein sample was used.
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Sample composition
- 20 mg/mL γ-globulin
- 3% Mannitol
- 2% Sucrose
- 10 mM L-arginine
- 0.01% Tween20
Concentration of spiked CHO gDNA
10 ng/mL
1 ng/mL
100 pg/mL
Detection Wavelength
| CHO gDNA | 520nm (e.g. FAM) |
| Internal Control | 554nm (e.g. HEX) |
Internal Control is non-natural synthetic DNA.
Overview / Applications
| Outline | This kit detects host cell-derived genomic DNA remaining in biological products such as vaccines and biopharmaceuticals by the qPCR methods using the TaqMan probe. Our unique polymerase configuration enable highly sensitive detection of trace amounts of DNA contained in samples. In order to suppress human error during liquid preparation and improve the work efficiency and the test accuracy, the PCR reaction solution is prepared in one bottle as 1 x PCR Master Mix. In addition, it contains the primers/probes and the templates for internal control, so you can confirm that the PCR reaction is performed correctly. When used in combination with the separately sold [DNA Extractor Kit], it is possible to consistently extract and detect genomic DNA in protein solution prepared using CHO cells. |
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| Purpose | Detection of residual DNA derived from the host cells, which is one of the quality control for biopharmaceuticals. |
| Application | [Preparation of CHO control DNA for standard] 1) Mix 10uL of CHO Control DNA included in the kit and 990uL of DNA Dilution Buffer included in the kit. 2) Dilute the CHO Control DNA diluted in step 1) 10-fold with DNA Dilution Buffer to prepare a dilution series of 0.003pg/uL to 300pg/uL. 3) Perform the PCR reaction according to the package insert. |
Property
| Specificity | Confirmed that amplification does not occur in human, E. coli, and yeast DNA. |
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Manufacturer Information
Alias
For research use or further manufacturing use only. Not for use in diagnostic procedures.
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