Other than our own sample extraction products, which can be found at www.omegabiotek.com, Omega Bioservices does not recommend any specific product or procedure for extraction and purifying DNA and RNA. Labs should use the methods already in place that successfully provide purified DNA and RNA for other molecular biology applications. If quality issues do arise, we are happy to consult to provide effective solutions for any sample type.
Please note that Omega Bioservices offers a complete line of sample extraction services for any clinical or environmental sample type. Because we understand that quality DNA/RNA in equals quality NGS data out, we prefer to take full control of sample quality from the beginning to the end of the NGS process. We offer very cost-effective pricing on our sample extraction services, especially for large NGS projects. Please contact us for more information.
Sample quality and quantity
If you choose to provide the purified DNA or RNA for your selected NGS service, we prefer that measurements of DNA concentration are taken with an intercalating dye and fluorimeter method such as Qubit (LifeTechnologies) or Quantus (Promega). Measurements of RNA concentration can be taken via A260/A280, or via fluorimeter with RiboGreen dye if available. If A260/A280 measurements, such as with the Nanodrop, are used for either DNA or RNA quantification, please be sure to send much more material than is indicated in the table below to account for the potential quantity overestimation which is common with this method.
The listed amounts of requested input material also include enough to accommodate our in-house QC testing needs. For RNA, this testing includes the determination of RIN score, or DV 200 as appropriate. If quality of received material is not sufficient to guarantee quality results, for example a RIN score below 7.0 for RNA, we will consult directly with the customer prior to proceeding with NGS service.
Please use the following table for sample submission guidelines:
|Next generation sequencing method||Input material needed||Minimum amount required||Minimum concentration required|
|Whole Exome Sequencing – Illumina||gDNA||100 ng||5 ng/ÂµL|
|Whole Exome Sequencing – Agilent||gDNA||1.5 Âµg||10 ng/ÂµL|
|Whole Genome Sequencing||gDNA||2 Âµg||5 ng/ÂµL|
|16S, PCR-Targeted Sequencing||gDNA||12.5 ng*||5 ng/ÂµL*|
|FFPE DNA||gDNA||500 ng||5 ng/ÂµL|
|FFPE RNA||Total RNA||100 ng||25 ng/ÂµL|
|Total RNA Seq||Total RNA||2 Âµg**||25 ng/ÂµL|
|mRNA Seq||Total RNA||1 Âµg**||25 ng/ÂµL|
|Small RNA Seq||Total RNA including small RNA fractions***||2 Âµg||25 ng/ÂµL|
*Optimal quantity. Some sample types, such as swabs, may not yield optimal amounts of DNA.
**Optimal quantity. We frequently and successfully work with much lower amounts of RNA.
***Ensure sample extraction method for total RNA preserves constituent small RNA population.
For limited sample material, we have strategies that can accomplish NGS with much smaller amounts of DNA or RNA. Please contact us to discuss if this is the case.
For RNA Seq applications, if a customer has already converted RNA to cDNA, then we only need 100 ng for NGS. If a customer is sending us pre-made libraries, we ask for a minimum volume of 10 ÂµL at a concentration of 20nM.
All NGS submissions should be suspended in nuclease-free water or standard elution buffer (10 mM Tris, pH 8.5). Some customers may be utilizing the newer sample stability technologies that eliminate the need for cold chain transport. We also have the capability to work with any of these sample matrices, but please let us know that this is the case before your sample arrives. If shipping internationally with potential delays expected, please contact us for non-cold chain transport solutions for sample stability during shipping.
DNA can be shipped in several form, depending on the customer needs. We have received and successfully worked with DNA shipped in the following manners: dry in filter columns, spotted on filter paper, suspended in water or Tris, and shipped at either ambient temperature, cold packs, ice, or dry ice.
RNA is best shipped on dry ice, however RNA may also be shipped at ambient temperature when suspended in our E.Z.N.A.Â® RNA Transport buffer. This is beneficial for our international customers that might anticipate a delay in the shipping process. Please contact your distributor for information, or it may be purchased (catalog number R0527-00) through our website.
Please include a sample submission form (provided by us) in all shipments. Samples should be clearly labeled, placed in organized boxes, sealed in a zip-lock plastic bag and shipped to:
Attention:Â Sample Receiving
400 Pinnacle Way, Suite 450
Norcross, GA 30071
Phone contact is: 770-931-8400