Certal Residual DNA Detection Kits
For sensitive probe-based detection of residual CHO host cell DNA, including an internal control
- Low limit of quantification with excellent reproducibility
- High linear range
- Reliable and precise internal control
- Optimized protocols for a broad range of real-time cyclers
- Complete kits including internal control and positive control
When working with biopharmaceuticals, it is essential to evaluate the final therapeutic material to ensure that it is free of contaminating substances, such as residual DNA from the host cell. Certal Residual DNA Detection Kits are designed for sensitive and rapid real-time PCR quantification of residual host cell genomic DNA using dual-labeled hydrolysis (also known as TaqMan®) probes. The Certal CHO Detection Kit contains reagents for duplex real-time PCR detection of CHO residual host cell DNA, plus the Certal Internal Control. The kit also includes the Certal Residual DNA Positive Control, which verifies the functionality of the PCR assay in the absence of a signal from CHO host cell DNA, and 2 formats of ROX passive reference dye in different concentrations, enabling use on virtually any real-time instrument.
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Certal CHO Detection Kit (100)
For 100 x 25 µl reactions: 1.25 ml Certal Residual DNA PCR Master Mix, Certal Internal Control DNA and Assay, CHO DNA Positive control and CHO-Assay, 100 µl Uracil-N-Glycosylase, 210 µl High-ROX Dye Solution, 210 µl ROX Dye Solution, Nucleic Acid Dilution Buffer, TE Buffer
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211822
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The Certal Residual DNA Detection Kits are intended for molecular biology applications in pharmaceutical product testing. These products are not intended for the diagnosis, prevention, or treatment of a disease.
See trademarks.
可靠检测QIAGEN内参。|Certal Residual DNA Detection Kit实验流程。|在Rotor-Gene Q实时荧光定量PCR仪上灵敏、可靠的检测CHO基因组DNA。|检测最低阈限。|在多种PCR仪上进行灵敏、可靠的检测。|高度灵敏、可靠的检测。|
在Certal ResDNA Master Mix中加入靶序列(CHO)的引物和探针,以及内参。内参模板和样本模板加入后,反应开始,可以在多种PCR仪上进行反应。也可以选择性检测阳性CHO对照,可使用UNG进行处理,消除PCR产物的残留污染。|在Certal ResDNA Master Mix中加入靶序列(CHO)的引物和探针,以及内参。内参模板和样本模板加入后,反应开始,可以在多种PCR仪上进行反应。也可以选择性检测阳性CHO对照,可使用UNG进行处理,消除PCR产物的残留污染。|图中曲线分布均匀,内参CT值稳定,说明在Rotor-Gene Q实时荧光定量PCR分析仪上使用Certal Residual DNA Assay检测DNA具有高度可靠性,也表明CHO基因组DNA的浓度没有影响对内参的灵敏检测。|检测阈限(LOD)是指重复24次检测都获得100%阳性扩增结果的最低DNA浓度,Certal CHO Detection Kit Assay的检测阈限是1 fg。定量检测阈限(LOQ)是指计算获得的CT值与CT标准值误差在±0.5以内的最低浓度,为10 fg。|在多种常用的PCR仪上使用Certal CHO Detection Kit检测起始量为3 fg至30 ng的DNA。检测结果符合线性规律,表示在不同的qPCR仪上,Certal试剂盒均能获得可靠结果。|在Applied Biosystems ABI 7900 HT(384孔板规格)上使用Certal CHO Detection Kit检测CHO DNA(0.3 fg至300 ng)。起始DNA量为10 fg至300 ng时,检测十分灵敏、阈限低,符合线性规律。LOQ: 定量检测阈限,是指计算获得的CT值与CT标准值误差在±0.5以内的最低浓度。这一数值为实验得出:未知样本的平均CT值与标准曲线的偏差不能大于0.5。LOD: 检测阈限,是指重复24次检测都获得100%阳性扩增结果的最低DNA浓度。|
原理
To ensure that contaminating substances from the host cell line used to produce biopharmaceuticals are absent from final therapeutic material, Certal Residual DNA Detection Kits deliver a predesigned assay to specifically detect CHO host cell DNA. Each Certal Residual DNA Assay consists of an optimized primer pair and a FAM labeled hydrolysis probe for reliable detection of residual CHO genomic DNA. Human background DNA and Certal Internal Control DNA will not interfere with the Certal Residual DNA Assay.
The premixed Certal Internal Control Assay contains a forward and reverse primer and a MAX labeled hydrolysis probe for the detection of Certal Internal Control DNA. It has been specifically optimized to prevent interference with Certal Residual DNA Assays. The Certal Residual DNA Positive Control enables the correct interpretation of negative results by testing the functionality of the amplification reaction itself.
Certal ResDNA PCR Buffer has been optimized to deliver sensitive and reliable quantitative, duplex real-time PCR results. The master mix also contains dUTP, enabling pretreatment with uracil-N-glycosylase (UNG) prior to starting PCR to ensure that any contaminating PCR products do not affect subsequent PCR reactions.
| Certal Residual DNA Assays and control DNA |
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| 50x Certal Residual DNA Assay |
Predesigned, optimized, and validated primer-probe set (FAM labeled) |
Reliable detection of residual CHO host-cell DNA |
| Certal Residual DNA Positive Control |
Positive control DNA that will be amplified by the Certal Residual DNA Assay |
Verification that the Certal Residual DNA Assay is working correctly in the case where no residual host-cell DNA is detected in the sample |
| 50x Certal Internal Control Assay |
Predesigned, optimized, and validated primer-probe set (MAX labeled) for Certal Internal Control DNA detection |
Specific detection of Certal Internal Control DNA only. Optimized to prevent interference with Certal Residual DNA Assays |
| Certal Internal Control DNA |
Internal Control DNA that will be amplified by the Certal Internal Control Assay |
Excludes the presence of PCR inhibitors or can be used to control purification and amplification when added during the purification step |
| Certal ResDNA PCR Master Mix |
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| HotStarTaq DNA Polymerase |
15 min activation at 95°C |
Set-up of qPCR reactions at room temperature |
| Certal ResDNA PCR Buffer |
A balanced combination of KCl and (NH4)2SO4 ions |
Specific primer annealing ensures reliable PCR results |
| Synthetic Factor MP |
Reliable multiplexing analysis of up to 4 genes in the same tube |
| dNTP mix |
Includes dUTP, which partially replaces dTTP and enables optional UNG treatment |
Effective elimination of carryover of PCR products when the reaction is pretreated with uracil-N-glycosylase (UNG) |
| Additional components |
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| 50x ROX Dye Solution |
Separate tube of passive reference dye for normalization of fluorescent signals on Applied Biosystems 7500 real-time PCR systems. Optional: Can be used on Stratagene instruments from Agilent |
Precise quantification on cyclers that require ROX dye. Does not interfere with PCR on any real-time cycler |
| 50x High-ROX Dye Solution |
Separate tube of passive reference dye for normalization of fluorescent signals on Applied Biosystems 7900 and StepOne real-time PCR systems |
| Uracil-N-Glycosylase (UNG) |
Separate tube of pure, nuclease-free recombinant enzyme |
Effective elimination of carryover of PCR products |
| QuantiTect Nucleic Acid Dilution Buffer |
Proprietary buffer formulation for dilution and storage of nucleic acid standards |
Stabilizes RNA and DNA standards during dilution and reaction setup and prevents loss of nucleic acids on plastic surfaces, such as tubes or pipet tips |
操作流程
Certal Residual DNA Detection Kits provide a simple procedure for the detection of host-cell DNA, the Certal Internal Control, and the Certal Residual DNA Positive Control (see figure Certal Residual DNA Detection Kit procedure). Simply mix the supplied master mix with the Certal Residual DNA Assay, the Certal Internal Control Assay and the Certal Internal Control DNA, and then add your sample DNA or Certal Residual DNA Positive Control. The kit handbook contains optimized protocols for use on a wide range of cyclers. ROX is supplied in 2 tubes of different concentrations, enabling use on any real-time instrument (see table "Choosing the correct ROX concentration").
| Rotor-Gene Q |
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| Rotor-Gene 6000 and Rotor-Gene 3000 |
X |
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| Applied Biosystems 7500, ABI ViiA7 |
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X |
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| Applied Biosystems 7900HT, StepOne and StepOne Plus, ABI PRISM 7700, 7000 and 7300 |
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X |
| Mx3000P, Mx3005P, Mx4000 |
X |
Optional |
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| iCycler iQ System |
X |
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| LightCycler 480 |
X |
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应用
Certal Residual DNA Detection Kits enable sensitive real-time PCR using sequence-specific probes for the detection of host-cell DNA, including an internal control. The kits can be used on a wide range of real-time cyclers, including cyclers from QIAGEN, Applied Biosystems, Bio-Rad, Roche, and Agilent.
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FAQ ID -2973
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FAQ ID -2974
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FAQ ID -2975
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FAQ ID -2976
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FAQ ID -2977
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FAQ ID -2978
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FAQ ID -2979
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FAQ ID -2980
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For sensitive real-time PCR detection of residual host cell DNA and an internal control using sequence-specific probes
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图片
可靠检测QIAGEN内参。
在Certal ResDNA Master Mix中加入靶序列(CHO)的引物和探针,以及内参。内参模板和样本模板加入后,反应开始,可以在多种PCR仪上进行反应。也可以选择性检测阳性CHO对照,可使用UNG进行处理,消除PCR产物的残留污染。
Certal Residual DNA Detection Kit实验流程。
在Certal ResDNA Master Mix中加入靶序列(CHO)的引物和探针,以及内参。内参模板和样本模板加入后,反应开始,可以在多种PCR仪上进行反应。也可以选择性检测阳性CHO对照,可使用UNG进行处理,消除PCR产物的残留污染。
在Rotor-Gene Q实时荧光定量PCR仪上灵敏、可靠的检测CHO基因组DNA。
图中曲线分布均匀,内参CT值稳定,说明在Rotor-Gene Q实时荧光定量PCR分析仪上使用Certal Residual DNA Assay检测DNA具有高度可靠性,也表明CHO基因组DNA的浓度没有影响对内参的灵敏检测。
检测最低阈限。
检测阈限(LOD)是指重复24次检测都获得100%阳性扩增结果的最低DNA浓度,Certal CHO Detection Kit Assay的检测阈限是1 fg。定量检测阈限(LOQ)是指计算获得的CT值与CT标准值误差在±0.5以内的最低浓度,为10 fg。
在多种PCR仪上进行灵敏、可靠的检测。
在多种常用的PCR仪上使用Certal CHO Detection Kit检测起始量为3 fg至30 ng的DNA。检测结果符合线性规律,表示在不同的qPCR仪上,Certal试剂盒均能获得可靠结果。
高度灵敏、可靠的检测。
在Applied Biosystems ABI 7900 HT(384孔板规格)上使用Certal CHO Detection Kit检测CHO DNA(0.3 fg至300 ng)。起始DNA量为10 fg至300 ng时,检测十分灵敏、阈限低,符合线性规律。LOQ: 定量检测阈限,是指计算获得的CT值与CT标准值误差在±0.5以内的最低浓度。这一数值为实验得出:未知样本的平均CT值与标准曲线的偏差不能大于0.5。LOD: 检测阈限,是指重复24次检测都获得100%阳性扩增结果的最低DNA浓度。
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