Sanger Sequencing Project and Sample Submission Requirements
Project Requirements
All requests for Sanger Sequencing must be submitted through the Sanger Sequencing LIMS. Please contact Kristen Pike for more information regarding submitting requests and costs.
Sample Requirements
The table below summarizes the required template concentrations for efficient and reliable processing of your samples. We recommend that samples be quantified via NanoDrop or PicoGreen prior to normalization in water and shipment to Laboratory of Molecular Technology.
| Description of Service | Template Type | Min. Vol. (uL) | Conc. (ng/uL) | Total DNA (ng) | Format |
| Amplicon Sequencing 100-200 bp |
Amplicon | 10 | 1-3 | 10-30 | Tube or Plate |
| Amplicon Sequencing 200-500 bp |
Amplicon | 10 | 3-10 | 30-100 | Tube or Plate |
| Amplicon Sequencing 500-1000 bp |
Amplicon | 10 | 5-20 | 50-200 | Tube or Plate |
| Amplicon Sequencing 1000-2000 bp |
Amplicon | 10 | 10-40 | 100-400 | Tube or Plate |
| Amplicon Sequencing >2000 bp |
Amplicon | 10 | 20-50 | 200-500 | Tube or Plate |
| Plasmid Sequencing | Plasmid | 10 | 30-40 | 300-400 | Tube or Plate |
| BAC Sequencing | Cosmid, BAC | 10 | 500- 1000 | 5,000-10,000 | Tube or Plate |
| BAC Sequencing | Bacterial gDNA | 10 | 2000-3000 | 20,000-30,000 | Tube or Plate |
Sequencing Primer Requirements
A list of commonly used sequencing primers supplied by the LMT can be found here. For primers supplied by the customer, a minimum of 10 uL of primer for each template at a concentration of 0.5 uM is required. For optimal results, we recommend a sequencing primer length of 10-20 bases long, with a melting temperature (Tm) of ~ 55ºC. Please see our High-throughput or Low-throughput Sequencing Sample Preparation Guidelines for more detailed information regarding sample and primer preparation requirements.
Shipping Information
Subsequent to placing a request for service in the Sanger Sequencing LIMS, please send your samples along with a copy of your LIMS generated request worksheet 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: LMT Sanger Sequencing Group
- Building and Room Number: ATRF, 8560 Progress Drive, Wing C, 3rd Floor
- Phone Number: (301)-846-5676
- Type of Shipment: DNA
