How to Submit DNA for Sequencing
- Initiate a Sanger Sequencing Order by completing this form.
- Download and fill out the Sequencing Samples Form linked in the email and send it to genomics@ospifse.net.
- DNA samples should be submitted in nuclease-free, sterile water. NOT in Elution Buffer or TE.
- Be sure to accurately quantify your samples (via Nanodrop, Qubit—both available in the facility).
- Recommended DNA concentrations for PCR samples:
- PCR Product Size | Concentration
- 100 – 200 bp | 1 – 3 ng/μL
- 200 – 500 bp | 3 – 10 ng/μL
- 500 – 1000 bp | 5 – 20 ng/μL
- 1000 – 2000 bp | 10 – 40 ng/μL
- >2000 bp | 20 – 50 ng/μL
- All PCR products must be cleaned (i.e. gel purification, Qiagen MinElute kit, or SPRI bead clean up) to remove excess primers, buffer, dNTPs and other contaminants that can inhibit the sequencing reaction
- The concentration of DNA for plasmid sequencing is determined as follows:
- (Plamid size in kbp) * (25 ng/μL)
- Submit at least 5 μL of your sample/rxn. Unused sample can be retrieved from the facility after you receive your sequence data.
- Be sure to write the appropriate requisition numbers on the tube caps for each DNA sample.
- Primers should be submitted with at least 5 μL/rxn at 3.2 μM.
- The facility is happy to provide standard sequencing primers (M13 for and rev, T7, T3 and Sp6) for no additional cost.
- Please be sure to include if the samples are AT or GC rich. (This affects the sequencing buffer we use.)
- Drop off samples in the freezer outside Porter 510 and place requisition sheet in the tray below the freezer.