- Service Description
- Experienced - MO BIO has over 22 years of experience in providing DNase-free certifications to many major medical device companies and suppliers of plastic consumables
- Qualified - All MO BIO Services technicians receive extensive training and are qualified to test raw materials or end products for the presence of DNase
A deoxyribonuclease, or DNase, is an enzyme degrades DNA by catalyzing the hydrolytic cleavage of phosphodiester linkages in the DNA backbone. DNase contamination can come from contact with human skin, and is often present in the lab environment. DNase contamination is of great concern in the medical device and pharmaceutical industries as well as the biotech and research fields, because DNase can cause degradation of valuable DNA samples, which may make it impossible to analyze the DNA via PCR, QPCR or next generation sequencing.
Certifying products DNase-free has become a common practice among general laboratory plastics manufacturers as well as medical device and pharmaceutical companies. Testing your product lines for this common contaminant will add integrity to your products and will enable a wider range of customers to utilize your products for their experiments and applications.
Figure 1. The effect of DNase contamination on DNA. Samples were incubated for 1 hour at 37°C, followed by inactivation for 5 minutes at 65°C. Results are shown on a 1% agarose gel. No degradation was observed in lanes 3-4, which did not contain DNase. Samples in lanes 1-2 were degraded due to the presence of DNase contamination. Lane 5 is a control and did not contain DNase.
DNase Test Method
All testing apparatus are treated with Diethyl pyrocarbonate (DEPC) to inhibit possible DNase contamination of the experiment from outside sources. Test samples of the product are extracted and a portion of extract is incubated with DNA standard (1 kb DNA ladder) in a buffered solution. Negative and positive controls are run along side the test sample extract to validate the experimental procedure and technique. An unexposed DNA standard is used as the negative control. A DNA standard is exposed to DNase is used to represent the positive control.
All samples are incubated 1 hour at 37°C. After the incubation, the reactions are evaluated by agarose gel electrophoresis and a picture for the Certification of Analysis and final report is taken. The test samples and the negative control should show no degradation. The positive control should be very smeared, indicating the effect of DNase contamination on the DNA standard pool that is used for all the reactions.
Passing test criteria: The DNA exposed to the test sample or extract must look exactly the same as the DNA negative control that was exposed to sterile water. The negative control must be completely intact with no degradation. The positive control must show degradation of the DNA standard for the test to be valid.
Test Sensitivity: 10-7 Kunitz Units/ µl
How To Begin Certifying Your Products DNase-Free
To begin certifying your product DNase Free, a sample of the product must be received by our Services Division to evaluate how best to extract your particular product for testing. Once an extraction procedure is approved, it is used each time that product is received for testing. Many products will fit one of our pre-designed extraction procedures, such as microcentrifuge tubes, PCR tubes, pipet tips and PCR plates. However, not all products are the same, thus all products are evaluated on an individual basis. Liquid solutions and reagents as well as solids, due to the diversity of these products, require a separate test procedure to be designed each time a product of this type is submitted.
- Contact Services
Our Services team has over 23 years of experience in providing services that keep your products competitive and in demand. Our customers include many of the top plastics and medical device suppliers in the industry. Our services specialists can also work with you to develop a custom testing schedule and/or custom tests that fit the specific needs of your products. We are located in Carlsbad, CA, USA.