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Sample collection related questions:
Mainly buccal/cheek cells
When collecting all the swabs supplied in the iSWAB kit you generate enough concentrated yields required for the majority of genomics downstream applications. However, based on our testing, collecting early in the day before brushing your teeth could give 15-20% extra yield.
Abstain from eating, chewing gum, smoking brushing teach or drinking 30 minutes before collecting the sample. Especially abstain from drinking hot beverages, such as coffee, tea, etc.
We recommend using all the swabs provided in the kit. This will ensure getting the highest yields.
The protocol calls for 20 seconds for each swab. While swabbing , try to rotate the swab in a full rotation o maximize the amount of buccal cells collected by each swab.
Because of the ease of use and collection method, the iSWAB is suitable for any age group from infants to elderly. It can be self collected or can be done by assisted collection by another house hold member, guardian or staff member.
After using all the swabs provided in the kit, the collected sample should be cloudy/milky with turbidity similar to the image below labeled with “Proper Collection”
If the sample is not cloudy/milky as described above, repeat the swabbing process after 2 hrs or next day in the morning using Q-tips swabs or ear wax cleaning tips or any standard cotton swabs. Usually using two extra swabs for the re-swabbing might be sufficient, but make sure the liquid in the tube looks similar to the image above before shipping the sample back to the processing lab.
The collected sample can be shipped under ambient temperature. No need for any refrigeration or cold chain transport involvement.
This depends on the yield requirement for your downstream application. You can prep 200-300µL or the whole sample volume in the iSWAB tube and leave the rest for future use. However, there are extraction methods that will allow you to process the content of the whole tube at once.
The insert inside the iSWAB tube is designed to allow for a standard 200-300 µL pipette tip to access the collected sample in the iSWAB tube. Some robotic tips, such as from the Hamiltons, Tecans, and the Bravo instruments have narrow neck and large access volumes as high as 500µL.
The iSWAB was designed to simplify the sample collection as well as the processing. The majority of automated liquid handling systems/robotics has predesigned racks for the iSWAB screw cap self-standing tube. Another option is to use Abgene 48 tube rack (AB-1434) or any other tube rack with SBS footprint. Also, the end user can dispense the content of the tubes into a 96 Well 2.2 mL deep well plate to help increase processing throughput.
Please check the data sheet for individual sample collection kits.
The proprietary iSWAB buffer is a unique formulation that not only stabilizes the samples at the point of collection, but performs a gentle lysis which slowly releases DNA from the cells into the buffer over a period of several hours. This feature enables the mixture to be used in a direct-to-PCR application, in which the extraction step is skipped entirely (check the resources section for further information) . The benefits of this application are:
- Time, cost, and labor savings by eliminating extraction reagents and processing time
- Rapid field-based analysis
For more complex downstream applications such as Next Generation Sequencing or microarrays, however, we still recommend performing a standard DNA extraction.
Unlike saliva collection methods, the iSWAB buffer is an alcohol-free preserving and lysis system. Therefore any commercially available gDNA extraction method for blood can be utilized with no further modifications to the protocol such as QIAamp, Gentra Puregene, Genfind v2 and DNAdvance (check the resources section for further information). Mawi also provides our own gDNA extraction kits in both filtration and mag bead-based chemistries. Please contact us at info@mawidna for more information.
Sample input guidelines:
Mawi Mini-Prep: 200-300µL
Gentra Puregene: 300µL to 1mL
Genfind v2: 300-600µL
Less than 1% per collected sample.
This information is proprietary and critical part of the iSWAB innovation.
One year based on real time stability testing and 5 years based on accelerated stability testing. Testing was performed based on ambient temperature storage and transport.
– Direct PCR applications
– DNA Sequencing: Sanger and NGS
– Genotyping: PCR and microarray based
– Gene expression
The iSWAB-DNA has been tested successfully with: ABI 3100, 3130, 3730XL, HISeq, MySeq, Ion Torrent, Proton, Illumina and Affymetrix microarray systems, Fluidigm, QuantStudio, and several other qPCR and PCR systems.
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