Our Sanger Sequencing lab uses an ABI 3130XL Automated DNA sequencer. We typically see read lengths of 600-800 bp. The turnaround time for this service is usually under two days and you will receive a text file and a chromatogram file with your sequence.
Both samples and primers can be submitted in separate tubes adjusted to the guidelines below:
Download the DNA Sequencing Order Form
Download the PCR SEQ Form
Plasmids:
- We recommend using Qiagen mini or maxi kits.
- Concentration should be determined by A260. Please adjust concentration to between 50ng/ul and 150ng/ul.
- Submit 300ng per sequencing reaction.
PCR Products:
- Gel purification may or may not be necessary depending on how clean the product is. Either way, the sample will need to be cleaned using a product such as Quiagen’s QIAquick kit.
- The concentration should be between 10ng/ul and 150ng/ul
- Submit 10-150ng per sequencing reaction depending on the length of the PCR product.
Primiers:
- Please adjust concentration to 5uM. This corresponds to 25ng/ul or 5 picomoles/ul.
- Submit 5ul per sequencing reaction.
We offer the following standard Primer Sequences free of charge. Simply indicate on the form which you would like us to use.
| T7: | 5′-TAA-TAC-GAC-TCA-CTA-TAG-GG-3′ | |
| T7 Term: | 5′-CTA-GTT-ATT-GCT-CAG-CGG-3′ | |
| T3: | 5′-ATT-ACC-CCT-CAC-TAA-AG-3′ | |
| M13F: | 5′-TGT-AAA-ACG-ACG-GCC-AGT-3′ | |
| M13R: | 5′-CAG-GAA-ACA-GCT-ATG-AC-3′ | |
| PGEX3: | 5′-CCG-GGA-GCT-GCA-TGT-GTC-AGA-GG-3′ | |
| PGEX5: | 5′-GGG-CTG-GCA-AGC-CAC-GTT-TGG-TG-3′ | |
| SP6: | 5′-ATT-TAG-GTG-ACA-CTA-TAG-A-3′ |
We sequence both PCR products and plasmid samples. See the appropriate form below.
Pricing:
| Tufts Customers: | $4 per reaction | ||
| External Customers: | $6 per reaction |