QIAcuity Residual DNA Quantification Kits

For checking carryover of host cell DNA with higher accuracy and precision using the QIAcuity Digital PCR System

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CHO resDNA Quant Standard Kit

Cat. No. / ID: 250223

CHO resDNA Quant Standard (1x), Rehydration Buffer
$924.00
Kit
resDNA Quant Standard
resDNA Quant
Cell type
CHO
HEK293
E. coli
QIAcuity Residual DNA Quantification Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

  • Detects even trace levels of contamination with high sensitivity
  • Offers a selection of HEK293, CHO and E. coli host cell DNA
  • Multicopy target assays unaffected by the fragmentation level of the resDNA

Product Details

Residual host cell DNA (HCD) monitoring is an important step in the process of manufacturing proteins and vaccines. The potential carryover of HCD poses a safety concern. Levels of HCD must not exceed those established by regulatory agencies such as the U.S. Food and Drug Administration and the World Health Organization.


Digital PCR is the choice of detection for residual DNA quantification due to its unmatched sensitivity and accuracy, especially at trace levels of contamination compared to the other detection methods such as qPCR.

Performance


The QIAcuity Residual DNA Quantification Kits provide accurate CHO, E. coli and HEK293 residual DNA (resDNA) quantification results even in the presence of PCR contaminants and other inhibitory reagents. Multicopy target assays ensure that results are unaffected by the fragmentation level of the resDNA, which is an issue when using qPCR.

Digital PCR provides higher sensitivity of detection at a lower template input range in comparison to qPCR and therefore enables a more robust application.

The kits work in conjunction with the QIAcuity Digital PCR System, QIAcuity UCP Probe PCR Kit and QIAcuity Nanoplates, offering an end-to-end fast dPCR workflow comparable to qPCR, but delivering an absolute quantification of resDNA in your sample. The kits have been designed with the requirements of bioprocess manufacturing and QC in mind.

Principle

The principle of the dPCR reaction in the nanoplates is described here.

Using digital PCR for host cell DNA monitoring improves the LOD/LOQ through partitioning of the bulk sample. Partitioning increases the effective concentration of the target, allowing small amount of host cell DNA to be captured and measured with higher accuracy and precision. The ability to load high template input amounts in the QIAcuity Nanoplate 26k combined with an increased number of partitions is one of the reasons for higher accuracy.

Procedure

The lysate is added to the residual quant assay together with the internal control and host cell DNA measured by absolute quantification. Copies/µl is converted to fg/µl via a provided conversion factor.

Applications

The QIAcuity Residual DNA Quantification Kits are ideal for the highly precise quantification of HCD in complex bioprocess intermediates.

FAQ

dPCR results are in copies per microliter. How do I calculate host cell DNA contamination in femtogram per microliter?

The conversion factors from copies per microliter to femtogram per microliter for each target assay (E. coli, CHO, and HEK293) will be provided in the quick-start protocol and the QIAcuity User Manual Extension: Application Guide.

FAQ ID - 3876
My samples are highly fragmented. Is the dPCR result for resDNA quantification accurate?

ResDNA Quant Kit (E. coli, CHO, and HEK293) assays are multicopy target assays that ensure quantification of  resDNA unaffected by the fragmentation level. In addition, short amplicon regions are selected for each target region. 

FAQ ID - 3877
Is DNA extraction from the samples needed?

No. DNA extraction is not required. However, if the samples contain high levels of PCR inhibitors or high protein concentration, DNA extraction is recommended for accurate quantification of ResDNA contamination.

FAQ ID - 3873
Is restriction enzyme digestion required?

Restriction digestion of the templates is required when the template length exceeds 20 kb for a better distribution of templates within partitions. Recommended restriction enzymes can be found in the applications guide and quick-start protocols.

FAQ ID - 3875
Which extraction kits are recommended?

QIAamp UCP DNA Micro Kits, QIAamp HMW MagAttract Kits, QIAsymhony DSP Kits, QIAsymphony Certal Kits, DNeasy Blood & Tissue Kits, and QIAprep Spin Maxi, Midi and, Miniprep Kits are few recommended extraction kits to combine with ResDNA Quant Kits.

FAQ ID - 3874
Is dPCR sensitive to inhibition?

dPCR is less prone to inhibition in comparison to qPCR; however, we recommend dilution of samples that are known to contain PCR inhibitors.

FAQ ID - 3880
How much sample can I load for detecting host cell contamination?

26k Nanoplates are recommended for resDNA quantification, which maximizes the amount of template that can be loaded within a single reaction. In 26k nanoplates, this corresponds to 28 μl in 40 μl reaction volume. In addition, maximum template loading amounts for each kit will be provided in QSPs. 

FAQ ID - 3879
What are the ResDNA Quant Kit components?

The kits are composed of a premixed master mix that contains the  target. The kit also contains an internal control assay detected in Green and Yellow channel, internal control DNA, positive control DNA, and dPCR qualified water. All necessary components are included in the ResDNA Quant Kits.

FAQ ID - 3878