sequencing - MetabolicEngineeringGroupCBMA/MetabolicEngineeringGroupCBMA.github.io GitHub Wiki
DNA sequencing
Recommendations from EUROFINS
| DNA | Size | Concentration | Volume |
|---|---|---|---|
| Plasmid DNA | - | 50-100 ng/µl | 15 µL |
| Purified PCR products | 150-300 bp | 2 ng/µl | 15 µl |
| Purified PCR products | 300-1000 bp | 5 ng/µl | 15 µl |
| Purified PCR products | 1000-3000 bp | 10 ng/µl | 15 µl |
| Unpurified PCR products | 150-300 bp | 4 ng/µl | 15 µl |
| Unpurified PCR products | 300-1000 bp | 10 ng/µl | 15 µl |
| Unpurified PCR products | 1000-3000 bp | 20 ng/µl | 15 µl |
Quantify the template concentration via agarose gel or a photometer to ensure accurate results.
- Put barcode stickers on all tubes
- Add 15 µl purified DNA with either of the concentrations given in the table above
- Add 2 µl of primer with a concentration of 10 pmol/µl (10 µM)
- The total volume of your premixed sample should be 17 µL
Recommendations from GATC
Sending your samples the right way Sample requirements: Take 5 µl template DNA with either of following concentration:
- Purified plasmid DNA: 80 - 100 ng/µl
- Purified PCR products:
- 150-300 bp: 2 ng/µl
- 300-1000 bp: 12 ng/µl
- 1000 bp - : 25 ng/µl
Add 2.5 µl of primer with a concentration of 10 µM (10 pmol/µl) Please send total sample amount of 10 µl in 1,5 ml tubes To ensure the best possible quality of sequence data, the total volume of the sample should not be less than 10 µl. We recommend measuring the DNA concentration on an agarose gel.