At DNA Genotek, we recently learned the details of study being conducted by Generation Scotland called The Scottish Family Health Study. We wanted to share some of this information with you here on The Genetic Link.
Study overview
Generation Scotland is creating an ethically sound, family and population-based resource to identify the genetic basis of common complex diseases. The Generation Scotland Scottish Family Health Study is a large-scale, long-term project, which aims to find out more about common illnesses, such as heart disease and stroke. It is known that heredity plays a role in these diseases. By collecting medical and genetic information from families, and by following their health over the coming years, Generation Scotland hopes to find out which lifestyle and inherited factors are connected with higher or lower risk of these illnesses.
Main challenges
The Generation Scotland Scottish Family Health Study aims to collect phenotype information and blood for biochemical analysis and DNA isolation from 25,000 people living in Scotland. As the name suggests, the Scottish Family Health Study is based on families, so they need at least one sibling and preferably other family members (aged 18 or over) as well. The inclusion of as many family members as possible, especially from large families, will increase the power of the study. However, for a variety of reasons, not all participants can make a clinic appointment. Collecting DNA samples from relatives who live in geographically dispersed locations may not be possible within a clinic setting. Others may be inconvenienced by a visit to the clinic. To overcome this barrier to compliance and to maximize the number of study participants, Oragene•DNA kits are mailed to such participants, with instructions for self-collection.
To read the full case study on this study and how Oragene•DNA helped overcome their recruitment challenge, please click here.
I hope you enjoyed learning more about this study. Be sure to subscribe to The Genetic Link so you don't miss any of our articles.
Genetic research into diseases and conditions that predominantly affect children has always been particularly challenging. Traditionally, when high quality and high quantity DNA is required for a research project (e.g. greater than several micrograms), blood has typically been the only option. Blood draws are painful for children, dreaded by parents and disliked by technicians who must collect from the frightened child. It is also much more difficult to draw blood from children than from adults, due to the relative size of their veins and their willingness to stay in a fixed position. Unlike adults, children often don't have the ability to reason with their fears. But when a lot of DNA is required, there have not been many options.
Readers of The Genetic Link know that Oragene•DNA for DNA from saliva is an ideal alternative to blood-based DNA collections but what if the target donor group is young children or infants or equally challenging, adults who are unable to spit?
It has always been our goal to provide high quality, non-invasive, DNA and RNA collection products and we now have a new product to address the needs of those unable to provide a sample by delivering saliva directly into the kit. The Oragene•DNA (OG-575) product offers a non-invasive, painless and convenient method to collect DNA from children and others who cannot spit.
The simple collection protocol can be found on the product brochure. The saliva collected with the absorbent sponges is squeezed (or pressed) into the proprietary V-notch in the collection funnel and flows into the tube. This process is repeated until 0.75 ml of saliva is collected. Sample collection with the OG-575 will produce a median yield of 17.3ug of DNA.This format is standardized for automated processing, increases efficiency, minimizes sample handling and reduces handling errors.
Oragene•DNA (OG-575) for assisted collection provides:
- Easy collection, transportation and processing
- Painless, non-invasive collection of high quality, high quantity DNA
- DNA that is stable for years at room temperature
- Compatibility with Oragene•DNA protocols
- DNA that is proven on downstream applications
- Tube and liquid sample that is compatible with automated workflow processes
We’re very excited about this new product and the feedback we’ve received so far has been extremely positive. If you’d like to request a trial kit of the Oragene•DNA (OG-575) product, please click here.
If you have any feedback on this product, you can leave a comment and let us know.
If you've been following our blog or our newsletter, you're probably aware that DNA Genotek has experienced significant growth in our partner program. We've added partners across all categories: genomics service providers, diagnostic service providers and technology vendors.
One of the newest partners to complete this validation process is Precision Biomarker Resources. Precision Biomarker provides automated, high-throughput microarray services to expedite the biologic investigations of pharmaceutical, biotechnology and academic researchers. Their services include: nucleic acid isolation, SNP genotyping, expression profiling, qPCR protocol design and processing, and custom bioinformatic analysis. Precision Biomarker Resources serves as a Genomics Service Provider for investigators working with DNA Genotek’s Oragene•DNA sample collection system.
Precison Biomarker is also the first partner to provide a special offer to Oragene•DNA customers to help them celebrate their successful completion of the partner program requirements. I'm pleased to share this news with you and pass along information about this limited time promotion.
From now until September 30th, 2010, Precision Biomarker Resources will include free DNA isolation from your Oragene•DNA samples as part of any whole genome genotyping project performed by Precision Biomarker Resources and using the Affymetrix SNP Array 5.0 or SNP Array 6.0 microarrays. A minimum of 30 samples must be submitted to qualify. Samples and a purchase order for the accompanying genotyping service must be received by Precision Biomarker Resources before September 30th, 2010.
To schedule your experiment now, please call Precision Biomarker toll free 888-857-0752 or e-mail sales@precisionbiomarker.com. Be sure to include your name, institution and contact information (phone number and e-mail) so that they can contact you if we need any additional information.
What do you think of DNA partners offering special incentives for Oragene•DNA customers? Would you like to see more offers like this? If so, leave a comment and let us know.
Photo credit: Alexandra Simms
A few weeks ago, I posted an article about best practices for long term storage of unpurified Oragene•DNA samples on The Genetic Link. The response to the article was positive so I'm following up with today's story about long term storage of DNA purified from Oragene•DNA samples.
The preservation and storage of DNA is an important consideration for molecular epidemiology and population studies. As the makers of the Oragene•DNA family of products, we have prepared these recommendations for the long-term storage of DNA purified from saliva samples collected with Oragene•DNA to help our customers achieve optimal results.
Preventing DNA Degradation
There are three major causes of DNA degradation in a purified sample (ref. 1). Samples may be accidentally contaminated by bacteria, but storage at 4°C or lower will minimize bacterial metabolism and the release of nucleases. DNases may be inadvertently introduced from the skin, but this can be minimized by wearing gloves when handling samples. Repeated cycles of freezing and thawing may also contribute to DNA degradation. This may be minimized by splitting the purified DNA into multiple aliquots and thawing one at a time.
There are two major causes of degradation of purified DNA:
- Acid hydrolysis - due to storage in a solution with no buffering capacity, explained in "Comparison of TE and water" section
- Nuclease activity - DNases may come from the skin of the person handling the sample, contaminated labware or bacterial contamination of the purified sample. Use of gloves can minimize contamination from the person and use of nuclease-free labware and working in a clean environment can prevent both nuclease and bacterial contamination. Additionally, inclusion of a metal ion-chelating agent, such as EDTA, in the storage buffer can effectively inhibit the activity of any DNases present in the sample.
Comparison of TE and water
Kasper and Lenz (ref. 2) performed an 8-year study of DNA stored in water or Buffer AE (10 mM TrisHCl; 0.5 mM EDTA, pH 9.0). DNA stored in Buffer AE at -20°C or 2-8°C showed no degradation by gel electrophoresis and amplified well in a PCR assay. DNA in water remained intact when stored at -20°C but samples were degraded when stored at 2-8°C and performed poorly in a PCR assay. Pure water lacks buffering capacity and an acidic pH may lead to DNA hydrolysis.
Biobank recommendations
An EU workshop on Biobanks (ref. 3) recommends freezing DNA samples to prevent bacterial contamination and to minimize evaporation of the sample. Tris-EDTA (TE) buffer contains sufficient buffering capacity to prevent acid hydrolysis of DNA. Similarly, the UK Biobank (ref. 4) recommends the storage of DNA in a nuclease-inhibiting environment at a temperature of -20°C, since no significant increase in stability is observed at temperatures below -20°C.
References
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O'Brien, D. (2002). High-throughput DNA purification. Modern Drug Discovery. 5(3), 25-26.
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Kasper, Y. et Lenz, C. (2004). Stable 8-year storage of DNA purified with the QIAamp DNA Blood Mini Kit. QIAGEN News. 2004 e10.
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Biobanks for health: Optimising the use of European biobanks and health registries for research relevant to public health and combating disease. Report and recommendations from an EU workshop held at Voksenåsen Hotel, Oslo. January 28-31, 2003.
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Sample handling and storage: Subgroup protocol and recommendations. Version 1.0. UK Biobank. July 7, 2004
We hope these best practices will help you optimize the long term storage of all your purified Oragene•DNA samples. If you like this article, be sure to let us know by leaving a comment.
Oragene•DNA is well known for a number of characteristics - ease of use, non-invasive collection, high quality and quantity DNA and, of course, long term storage at ambient temperature. It's often difficult for researchers and clinicians to believe that storage at ambient temperature is possible for DNA samples. We are frequently asked about this specification of our product. However, studies using Oragene•DNA prove that customers can rely on the ability to store Oragene•DNA/saliva samples at ambient temperatures when collecting samples in remote locations, via the mail, or for event-based recruitment. The figure to the right shows an agarose gel electrophoresis of DNA extracted from Oragene•DNA/saliva samples stored at room temperature for 5 years.
The preservation and storage of DNA samples is an important consideration for molecular epidemiology, clinical trials, and population studies. To ensure your success, we have created the following best practices for the long term storage of unpurified saliva samples collected with the Oragene•DNA Self-Collection Kit. We will also highlight the rationale for these recommendations.
Storage at room temperature
DNA from saliva is stable in the Oragene•DNA collection tube for up to 5 years. This stability is achieved as a result of the kit's proprietary reagents that inactivate enzymes in saliva and minimize chemical hydrolysis of DNA. These reagents also prevent the growth of bacteria in the sample.
Frozen storage
Oragene•DNA/saliva samples may also be stored at temperatures below -20°C. Samples may undergo at least three freeze-thaw cycles with no evidence of DNA degradation. Although the Oragene•DNA collection tube is designed to ensure a tight seal, frozen storage may further reduce evaporation of the liquid medium during long term storage.
Aliquots in microcentrifuge tubes
The Oragene•DNA collection tube is designed for user friendly saliva collection from donors. However, in the lab, storage in more compact tubes may be preferable. To reduce storage space, an Oragene•DNA/saliva sample which has been heated at 50°C for a minimum of 1 hour may be split into 4 aliquots of 1 mL each and stored in 1.5 mL screw-top microcentrifuge tubes. It is advisable to use tubes with O-rings to ensure a tight seal and minimize long term loss from evaporation.
We hope these best practices will help you optimize the long term storage of all your unpurified Oragene•DNA samples. If you like this article, watch for an upcoming blog post on the best practices for long term storage of purified DNA from Oragene•DNA samples. If you want to be notified of our blog posts, be sure to subscribe to The Genetic Link.
Was this article useful? Let us know what you think by leaving a comment.
Today, DNA Genotek announced our involvement in a significant pilot project with the UK's largest bone marrow registry, the Anthony Nolan Trust. The details of this exciting project follow:
DNA Genotek, a leading provider of products for biological sample collection, stabilization and preparation, today announced that The Anthony Nolan Trust, the UK's largest bone marrow donor registry, has selected Oragene•DNA for a pilot project aimed at increasing donor recruitment. Bone marrow donor registries, also known as HLA registries, use HLA DNA testing to match leukemia patients with prospective donors. The pilot project will determine if donor recruitment can be increased significantly with the use of non-invasive, saliva-based DNA collection compared to blood collection.
The Anthony Nolan Trust has traditionally required all registry participants to have a blood sample collected either at their doctor's office or by phlebotomists at recruitment clinics. Under this pilot project, the potential donor provides a saliva sample without any assistance (for example, at home) and mails the sample back through the regular postal system, thereby facilitating and dramatically reducing the cost of the entire process. DNA samples collected with Oragene•DNA are easier, safer, and faster to collect than other methods and remain stable at ambient temperature for transport and storage prior to being analyzed at the lab.
There are currently over 400,000 people included in The Anthony Nolan Trust's registry, but many more participants are needed. 70% of patients needing a transplant cannot find a compatible match from within their families and rely on registries to find an unrelated donor for life-saving procedures. Through using Oragene•DNA, The Anthony Nolan Trust expects an increase in donor recruitment and therefore the likelihood of matches between those patients needing bone marrow transplants and potential donors held in their database.
Ailsa Ogilvie, director of operations at The Anthony Nolan Trust stated: "We have historically recruited new people to our register using blood samples, but our early laboratory results confirm that Oragene•DNA provides the high quality DNA that we need. We are optimistic that this pilot project will simplify the recruitment process and ultimately increase the number of people willing to join our register. Following the successful completion of this pilot phase, we hope to switch completely to using saliva samples to recruit new donors to our register via Oragene•DNA from late 2010."
"By streamlining the sample collection and transportation procedures, Oragene•DNA enables The Anthony Nolan Trust to focus on its primary goal of increasing donor recruitment", said Ian Curry, president and CEO, DNA Genotek, Inc. "Oragene•DNA provides a reliable, cost-effective, and scalable method that is ideal for bone marrow donor registries worldwide. Our product is easy to use, safe to transport through the mail and remains stable at ambient temperature for long periods of time. What we learn from this pilot project will help chart the future for The Anthony Nolan Trust and we are certainly proud to be part of this effort."
Note: The pilot project referenced in this press release is currently available in limited areas in the United Kingdom. It is not expected to be available nationally until the summer of 2010.
Learn more at the European Immunogenetics and Histocompatibility Conference (EFI) in Florence, Italy May 17th when Alasdair J. McWhinnie of the Anthony Nolan Trust will present a session titled: "Saliva specimens collected with Oragene are a reliable alternative to blood and buccal swabs for large scale DNA extraction and HLA typing of recruits for hematopoietic stem cell donor registries." Register here.
The Genetic Link welcomes guest blog authors. Our first guest submission is from Rachel Dvoskin, Ph.D., with the University of Florida. We thank Rachel for her submission and welcome her as a guest author. I hope you enjoy this article.
At the department of Anthropology and Genetics Institute at the University of Florida, we study genetic variation in modern human populations to answer diverse questions ranging from the route early humans took when they first migrated out of Africa to the underlying causes of racial differences in susceptibility to complex diseases. To explore these varied aspects of human evolutionary history, we spend a lot of time figuring out how to collect DNA from a large number of volunteers. With today's genetic technology, all that's necessary is to obtain a small blood or saliva sample from each of our participants -- a task relatively easy to do in concept but quite a bit more challenging in practice.
When doing this kind of research -- whether at home or abroad -- we come up against a bunch of potential hurdles. Language and cultural barriers can make it difficult to explain what we are doing and what we are hoping to gain from our participants. As a blond, female, American anthropologist who could hardly say hello in Arabic, Dr. Connie Mulligan never expected collecting DNA samples to be as easy as it was in the remote corners of the Middle Eastern country of Yemen. The Oragene•DNA saliva collection kits are non-invasive, simple to use, and even include illustrated instructions right on the package, making their use completely self-explanatory. We had no problem collecting saliva samples from hundreds of Yemeni participants, including many women garbed in the full abaya.
Furthermore, the lack of need for freezing or refrigeration allowed us to travel around the country without worrying about storage conditions. The disks are small and self-contained, and we did not have to carry any separate buffers or processing equipment. The saliva collectors made it possible to obtain large volumes of high-quality DNA. This DNA is used to answer questions about the movement of early humans out of Arabia. We will soon be able to thank our Yemeni study participants by telling them something new about their country's role in the peopling of the world.
We are now gearing up to do a study here in our own backyard in Florida. In collaboration with cultural anthropologists, we are studying how genetic factors and the social environment interact to influence a person's susceptibility to hypertension. Because we are interested in racial disparities in health in the U.S., we chose to conduct our community-based study in Tallahassee, a city with a long history of racial tension and civil rights activism. Given the potential for misuse of genetic information and the rare but horrific cases of unethical medical experimentation on oppressed minorities, it is understandable that people would be fearful or reluctant to participate in a study of this kind. We expect, however, that the non-threatening nature of the Oragene•DNA saliva collection kits will put people at ease and increase participation.
We value highly this simple, non-invasive technique because unlike many studies in which researchers opportunistically collect samples from people who are already being treated in a hospital or clinic, we are instead approaching regular people in their homes -- trying to get a representative sample of all types of people within a community. We think that the more benign and user-friendly the collection techniques, the more useful they will be for our research and for future work with minority groups and previously unsampled populations. This includes people in regions where lab or storage facilities are scarce as well as populations that have been overlooked or underrepresented in medical and health research.
Did you like this guest authored article? Leave a comment and let us know.
For the second year in a row, DNA Genotek collected DNA samples onsite at the National Walk for Epilepsy on March 27th, 2010 in Washington, DC. Our participation in this event supports the efforts of the Columbia University Family Studies in Epilepsy Program and their study which is designed to identify genes that play a role in causing epilepsy. While at the Epilepsy Walk, I had the opportunity to interview Janine Rose, Research Associate at Columbia University about their epilepsy research and how Oragene•DNA has helped their study. You can view the recorded interview here or read the transcript below.
Shauna White, DNA Genotek: I'm here with Janine Rose with the Columbia University Family Studies in Epilepsy and we're here at the National Walk for Epilepsy in Washington DC in March of 2010. Janine, thanks for joining us today, it's great to have you here.
Janine Rose, Research Associate, Columbia University Family Studies in Epilepsy: Thanks for having us.
Shauna: Can you tell me about the research study you're working on and what you hope to achieve with it?
Janine: Yes. We are a genetic research program at Columbia University and what we're looking to discover are the genes that affect certain types of epilepsies, childhood epilepsies, specifically idiopathic generalized epilepsy and idiopathic generalized epilepsy is a type of epilepsy that doesn't have an origin, a set origin. So we are looking to recruit subjects for this particular kind of epilepsy and by DNA saliva, we look at what these genes do and further identify in hopes that further down the road, finding a cure or new therapy for these patients.
Shauna: Why did you decide to collect DNA with saliva samples?
Janine: For many reasons. Before, and that was before my time (with the study) they would collect blood as you know it is less likely to find subjects willing to go through that donation. It is very difficult to store, it needs to be refrigerated, there can be pain, there are very many reasons, it can be messy. I think that the saliva is small and portable and light and extremely practical - a super easy way to collect and people are delighted to participate and enchanted to look at how they are made and it's a great conversation piece. It breaks the ice every time so we are very happy with the product. We use it very willingly.
Shauna: This brings me to our work here this week at the National Walk for Epilepsy. When you heard about this event, you obviously saw a way to help your study. What did you think about and why did you get involved?
Janine: Well, I was involved the first time last year which I participated with Columbia University Family Studies. The year previous I came as a participant and I noticed at the event how many sponsors they had and I thought well, why not us? I thought that as direct sponsor, I would like for us to be involved because I saw a real number of people who would definitely embrace this project and people that would either be willing to participate or donate in some way and support what we are doing. And finding people is what this is all about. At the moment we use mostly physician referrals to find our subjects so I thought why not come directly here and show them what we're doing at Columbia. Even if they're not qualified to join the study, if they don't have epilepsy, they could at least be an unaffected control which is what we're hoping to collect today. If we get the funding to go on with the study, then further down the road it's our goal to include other types of seizures if we get funding to do so.
Shauna: How has DNA Genotek helped you with this project?
Janine: I have to say that we wouldn't be here today if it weren't for DNA Genotek. We are very very grateful for the help that you've provided us. First of all, getting the booth, getting the space, helping us organize the event. It's been a real sharing experience for both of us. I think it's a very good fit. People see exactly how we get the DNA and having them (DNA Genotek) personally on hand here on the spot to have them answer any questions they may have about the product or project is great. We're very very pleased.
Shauna: Tell me a little about the future of the study. When do you think you'll have some results? When will people know the outcome of this study?
Janine: It's very difficult to predict - to put a time period on such a momentus study because every time you find something new it takes you in a new direction. With our funding, it allows us to go so far. As more funding comes along, we hope to increase our study. Our investigation is about how epilepsy comes. Of course we're working to point very specifically at what can be done at the appropriate times. For the moment, we're still at the investigation stage. But we found a true confirmation of genetic information. We feel that is a very good study both for reasons of our specificity. We are very strict about our criteria - our inclusion criteria. This allows us to know exactly what we're looking at. We don't cloud it with other things.
Shauna: Thank you so much for taking the time today. Best of luck with the National Walk for Epilepsy and best of luck with the study.
Janine: Thank you, thank you for having us.
Would you like to see more articles like this one? Leave us a comment and let us know.
Today, DNA Genotek announced that our Oragene•DNA self collection kit (OG-500), has been selected as a winner in the 2010 Medical Design Excellence Awards (MDEA) competition, the premier awards program for the medical technology community.
As an all-in-one system for the collection, stabilization, transportation and purification of DNA from saliva, our flagship Oragene•DNA product has been rapidly gaining acceptance as the preferred method for non-invasive DNA collection by research institutions and commercial organizations around the world. Oragene•DNA enables our customers' research and genetic analysis programs with an aim to advance health research and healthcare delivery.
Our status as a winner of the prestigious Medical Design Excellence Awards competition highlights our commitment to excellence in product design and ease of use. MDEA is the only awards program that exclusively recognizes contributions and advances in the design of medical products. A comprehensive review of the entries was performed by an impartial, multidisciplinary panel of third-party jurors with expertise in biomedical engineering, human factors, industrial design, medicine and diagnostics.
I am proud to share this award with our product development partner, DW Product Development Inc. and with our dedicated employees. Their outstanding focus on quality for all aspects of our business is what makes awards like this possible.
Interested in trying this product? Click here for an evaluation kit.
Saliva is one of the most accessible of our body's bio-fluids making saliva sample collection easy and non-invasive. Saliva also harbours a wide spectrum of genetic data that can be used for genetic research and clinical diagnostic applications. It might surprise you to know that much confusion surrounds the source of genomic DNA in saliva. It certainly came as a surprise to me when I met with a number of customers on a recent trip across the continent.
In recent years, we've seen a marked increase in the desire to understand the genetic basis of disease as a means to improve patient diagnosis and treatment. While the range of research spans the spectrum of known diseases, all genetic research projects share one vital building block in that they require DNA as a starting point. Traditionally, DNA has been extracted from white blood cells extracted from whole blood. White blood cells are an excellent source of large amounts of high quality genomic DNA. However, because of the invasiveness and cost of obtaining, transporting and processing blood samples, researchers and clinicians have long searched for alternative methods. Over the past few years, saliva has become recognized as a very important and reliable alternative to blood samples for genetic research, clinical diagnostics, pharmacogenomics and more. What exactly is it that makes saliva such a good alternative to blood for genetic applications? It all comes down to the source of DNA in saliva.
Surprisingly, many people I spoke with assumed the source of DNA in saliva is strictly buccal epithelial cells, however, studies show that up to 74%(1) of the DNA in saliva comes from white blood cells. Yielding virtually the same amount of DNA per volume and the same DNA quality, saliva can be considered as good and as reliable a source of DNA for a wide variety of genetic applications. One thing to note however is that not all oral samples are equal.
Oral Sample Collection Methods
There are three methods for collecting oral DNA samples - dry, wet and non-invasive procedures.
Dry procedures require the donor to insert a cytobrush, buccal swabs or other collection device into the mouth where tissue is scraped from the gum and cheek surfaces. These methods collect primarily buccal cells that are lower quality and are potentially contaminated with bacteria from the teeth and other surfaces.
Wet procedures include swishing liquids in the mouth and spitting them into a collecting vessel. Mouthwash, which can contain a high percentage of alcohol content, is typically used for this procedure. The protocol, which can request the donor to swish for up to one minute, can burn and be uncomfortable for the donor. Mouthwash is also designed to remove bacteria from teeth and other mouth surfaces which results in a high amount of bacterial content being released into the sample.
Both the dry and wet methods described above do not prevent bacteria from growing in the sample and do not actively stabilize DNA. These methods also involve the insertion of an object or substance into the mouth. While it is arguably less invasive than venipuncture, it does not quite meet the definition of ‘non-invasive'.
Completely non-invasive collection using Oragene•DNA is a simple, painless procedure that requires the donor to spit into a collection device. After providing a sample and closing the device, a solution is released from the cap to mix with the saliva. This solution stabilizes the DNA for long-term storage at room temperature and prevents bacterial contamination. There is no requirement to insert an object or substance into the mouth - the user simply holds the tube and spits into it. In my opinion, that's the true definition of non-invasive DNA collection. As we like to say at DNA Genotek, "Just Spit, That's It!"
The high quality and high quantity of DNA collected with Oragene•DNA provides an excellent option for applications ranging from the largest molecular epidemiologic studies in remote locations, to bio-banking or HLA typing and even a single personalized medicine test.
Did anything in this article surprise you? Leave us a comment and let us know.
Interested in learning more about non-invasive DNA sample collection with Oragene•DNA? Register for our live webinar on April 21, 2010 at 11:00am ET.
References: (1) Thiede, C. et al. Buccal swabs but not mouthwash samples can be used to obtain pretransplant DNA fingerprints from recipients of allogeneic bone marrow transplant. (2000). Bone Marrow Transplantation. 25(5): 575-577.)