Sanger Sequencing
Sample Submission Guidelines
DNA Sequencing Sample Submission Guidelines
For the best Sanger sequencing results GENEWIZ strongly recommends you follow our Sample Submission Guidelines as closely as possible. If you have any questions, our Technical Support team is here to help!
(EU) Email: sanger.europe@azenta.com | Phone +49 (0)341 520 122-41
(UK) Email: genomics.service.uk@azenta.com | Phone +44 (0) 1279 873837
Sample Preparation
Follow our barcode labeling instructions to avoid delays in sample processing and data delivery:
For orders with templates and primers in tubes, please use 1.5 ml flip cap reaction tubes (no screw caps). Label your tubes with barcodes according to your selected sample type to facilitate sample alignment and automated processing. Please note we do not accept samples labeled with fluorescent or other dyes.
Dilute your custom sequencing primer to 10 or 5 µM (pmol/µl) according to the service type (Pre-Mixed, Pre Defined) using water. Please refer to the table below for minimum volume for each sequencing reaction.
For the amount of template required in each of our DNA Sequencing Services (Pre-Mixed, Pre-Defined), please refer to the tables below. Please make dilutions in water or Tris. For best results, do not use Tris-EDTA (TE) because EDTA will inhibit the sequencing reaction.
Pre-Mixed Tube or Plate/ Sanger-EZ/ Vario Plate
For the Pre-Mixed Service, prepare purified template (DNA) and primer in a total volume of 10µl in the same tube according to the table below.
| Template | Primer | Template + Your Primer | ||||
| DNA Type | Submitted in | Concentration | Volume | Concentration | Volume* |
Pre-Mixed Total Volume |
| Plasmids purified | 1.5 ml reaction tube |
30-100 ng/µl | 5 µl | 5 pmol/µl (µM) |
5 µl |
10 µl |
| PCR fragments purified: <500 bp 500-1000 bp 1000-3000 bp |
5 ng/µl 10 ng/µl 25 ng/µl |
|||||
| Plasmid purified | 96-well plate | 30-100 ng/µl | 5 µl | 5 pmol/µl (µM) | 5 µl | 10 µl |
| PCR fragments purified: <500 bp 500-1000 bp 1000-3000 bp |
5 ng/µl 10 ng/µl 25 ng/µl |
|||||
Pre-Defined Tube
For Pre-Defined Services, provide template (20 µl) and your primer (20 µl) in separate tubes for up to eight reactions according to the table below.
| Template | Primer | |||||
| DNA Type | Submitted in | Concentration | Volume | Submitted in | Concentration | Volume |
| Plasmid purified | 1.5 ml reaction tube | 30-100 ng/µl | 20 µl | 1.5 ml reaction tubes | 10 pmol/µl (µM) | 20 µl |
| PCR fragments purified: <500 bp 500-1000 bp 1000-3000 bp |
5 ng/µl 10 ng/µl 25 ng/µl |
20 µl | ||||
Pre-Defined Plate or Vario Plate
For Pre-Defined Sanger Sequencing service in 96-well plates, provide 20 µl template per well and for primer provide separately 100 µl per plate.
| Template | Primer | |||||
| DNA Type | Submitted in | Concentration | Volume | Submitted in | Concentration | Your Primer Volume Per Plate |
| Plasmid (purified) | 96-well plate | 30-100 ng/µl | 20 µl | 1.5 ml reaction tubes |
10 pmol/µl (µM) |
20 µl – for up to 8 reactions (Vario plate) 100 µl – for more than 8 reactions or full plate |
| PCR product (purified / unpurified) <500 bp 500-1000 bp 1000-3000 bp |
5 ng/µl 10 ng/µl 25 ng/µl |
20 µl | ||||
| Low concentration plasmid (EU) | 10 ng/µl | 20 µl | ||||
