Sample and Documentation Requirements for the submission of DNA samples for 16S analysis

For total DNA, use any conventional method or commercially available DNA isolation kits to extract DNA. Treat with RNase to rid RNA contamination from DNA sample. Be sure to include a Completed Total DNA Sample Submission Form with each sample submission.

For sequencing-ready PCR amplified DNA, please specify (on the sample submission form) the PCR primer sequences, the Illumina adapter sequences and the index sequences used during PCR. Use any commercially available PCR clean-up kit. Be sure to include a Completed PCR- Amplified Sample Submission Form with each sample submission.

If less DNA is available than the quantity specified below, please contact us to discuss alternatives. If the quality is poorer than specified below, please contact us to discuss alternatives. All DNA should be resuspended in nuclease-free water or 10mM Tris-HCl pH 8.5 buffer.

Total DNA Sample
Sample Quality: A260/280 ≥1.8 and a gel run with high molecular weight ladder showing little to no DNA sample degradation and no RNA contamination.

Sample Documentation: Provide printout of 260/280 result and picture of gel with clearly labeled ladder and sample(s).

Sample Quantity: >100 ng at ≥ 10ng/uL unless arranged otherwise on Service Agreement.

Sequencing-ready PCR Amplified DNA Sample
Sample Quality: A single product at ~the expected size when analyzed on the DNA1000 chip of the Agilent 2100 Bioanalyzer. There should be no other peaks except the markers at 15 and 1500bp. All 13 points of the ladder should be distinctly identifiable.

Sample Documentation: Provide pdf printout of bionanalyzer results of the final Prepared Library(s). Provide sequences of adaptors and PCR primers and index primer sequence used (if any). The Prepared Library(s) must be compatible with sequencing primers and flowcell provided by Illumina.

Sample Quantity: 30 uL at ≥ 10nM unless arranged otherwise on Service Agreement

 

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750 Alfred Nobel Drive | Hercules, CA 94547 | Phone: (510) 964-0461 | Fax: (510) 964-9705 | Email: contact@eurekagenomics.com