Pyrosequencing Project and Sample Submission Requirements
Project Requirements
All requests for scientific and analytical work must be submitted through CSAS. Please contact Kristen Pike for more information regarding submitting requests, costs, and additional sources of funding for your project.
Sample Requirements
The table below summarizes the minimum requirements for assay processing. We recommend that samples be quantified via NanoDrop or PicoGreen prior to normalization and shipment to Laboratory of Molecular Technology. Since efficient and reliable processing of your samples is important to us, we would appreciate if you could send us more volume and a higher concentration of sample as your specimen source allows. This will enable proper sample QC and ensure that enough material is present even if evaporation occurs during shipment, or if it becomes necessary to repeat a reaction.
| Description of Service | Sample Type | Min.Vol.(uL) | Min. Conc. (ng/uL) | Total DNA (ng) | Format | Comments | |
| Optimizing PCR conditions per amplicon for snp genotyping (non-human) | gDNA | 10 | 10 | 100 | Tube or Plate | Required DNA amount is per amplicon to be assayed. Please provide control DNA . Required product amount is per Pyrosequencing assay run of 50-60 bp within a 250 bp PCR product. | |
| PCR & Pyrosequencing of gDNA for SNP detection (human) | gDNA | 5 | 10 | 50 | Tube or Plate | For optimization, commercial DNA will be used. Required product amount is per Pyrosequencing assay run of 50-60 bp within a 250 bp PCR product. | |
| Bisulfite Conversion of gDNA for % methylation detection by Pyrosequencing | gDNA | 10 | 10 | 250 | Tube or Plate |
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| Bisulfite-converted DNA (non-human) | Converted DNA | 15 | NA | NA | Tube or Plate |
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| Bisulfite-converted DNA (human) | Converted DNA | 2 | 10 | 20 | Tube or Plate |
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| PCR product that is run-ready for the pyrosequencer | PCR product | 10 | 10 | 100 | Tube or Plate | Required product amount is per Pyrosequencing assay run of 50-60 bp within a 250 bp PCR product. |
Subsequent to CSAS approval, please send samples to the Laboratory of Molecular Technology through the NCI-Fredrick courier. To schedule pickup, call transportation at (301)-846-1175 and provide them with the pickup location and the following drop-off information:
- Contact Name: Kristen Pike
- Building and Room Number: ATRF, 8560 Progress Drive, Wing C, 3rd Floor
- Phone Number: (301)-846-5676
- Type of Shipment: gDNA
