The TubeSeq Service is sequencing with the extra bit
Combined power of automation and expertise.
The Eurofins Genomics' TubeSeq Service combines long term expertise in automated Sanger sequencing processes with the handling of samples, which require bespoke solutions.
Highlights of the TubeSeq Service include:
- Order and re-order up to 8 reactions per sample
- Sequencing reactions can be paid with our TubeSeq Labels
- All types of sample material, incl. large constructs, are possible
- Option for Power Read Upgrade* for complex sequences
- Standard DNA reads are available next day after sample receipt
The benefits of our TubeSeq Service at a glance:
- High quality read lengths of up to 1,100 bases
- Free Q20, Q30 or Q40 monitored single strand data available
- DNA preparation and PCR purification is available
- Concentration adjustment for DNA samples can be selected
- Advanced analysis options are available, for free
- Different sequencing result files can be selected per order
- Secure online archive of sequence data for 100 days
- Free one time repeat, in case of technical failure
- Storage of template DNA and ordered primers for 4 weeks
- Storage of ordered primers for one year, on request
- Easy re-order options from stored DNA samples
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Sending samples the right way for our TubeSeq Service
Sending samples according to the requirements below helps us to do our job better and provides you with accurate results.
Sample submission
- Use 1.5 ml safe-lock tubes for your templates and primers
- Do not tape or wrap tubes with parafilm. Safe-lock tubes offer perfect sealing and evaporation protection
- Label your template and primer tubes with our barcode labels. TubeSeq Labels or Tube Labels
- Use a water resistant marker for any additional labeling of your tubes
Sample preparation
Use the following concentration and volume for your samples:
|
Sample type
|
Product length
|
Sample conc.
|
Vol. 1-4 rct.
|
Vol. 5-8 rct.
|
| Plasmid DNA |
< 30 kbp |
50-100 ng/µl |
15 µl |
20 µl |
| BAC/PAC/Cosmid DNA |
30 - 200 kbp |
200-1000 ng/µl |
15 µl |
20 µl |
| Purified PCR Products |
150-300 bp |
1 ng/µl |
15 µl |
20 µl |
| |
300-1000 bp |
5 ng/µl |
15 µl |
20 µl |
| |
1000-3000 bp |
10 ng/µl |
15 µl |
20 µl |
| Unpurified PCR Products |
150-300 bp |
4 ng/µl |
15 µl |
20 µl |
| |
300-1000 bp |
10 ng/µl |
15 µl |
20 µl |
| |
1000-3000 bp |
20 ng/µl |
15 µl |
20 µl |
Premixed samples (a mixture of template and primer):
- Templates should consist of 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)
- Ensure that the total volume of your premixed sample is 17 µl
Sequencing primers
Optimum primer conditions:
- Primers must not contain phosphorylation or fluorescent dyes
- The optimum primer length is between 16-25 bases
- The primer melting temperature (Tm) should be 50 - 62°C
- The GC content of the primer should be 35-60%
- Ideally one G or C should be located at the 3' primer end
- The number of 3' Gs or Cs should not exceed 2 Gs or Cs
- If possible, avoid >3 identical bases in a row in the sequence
Primer concentration and volume:
- Exactly 10 pmol/µl primer concentration is required per sequencing reaction
- Each primer must have a total volume of 15 µl (double distilled water or 5mM Tris-HCl); 5 µl of primer volume is required for every additional sequencing reaction
- Concentration of primers with wobble bases must be calculated according to the following formula: nX x ConcPrimer