LightRun Services

Sample requirements:

Take 5 µl of the purified template DNA with either of following concentration:

• Purified plasmid DNA:
  50 - 100 ng/µl
• Purified PCR products:
  150-300 bp:    1 ng/µl
  300-1000 bp:   5 ng/µl
  1000-3000 bp: 10 ng/µl

Add 5 µl of primer with a concentration of 5 pmol/µl (5 µM)

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.

Sample Submission:

Condition:

DNA templates must be purified. Sequencing reactions cannot be repeated and all reactions will be charged. Samples are not stored. For samples containing difficult-to-read streches (GC-rich, hairpin structurs...) we recommend our TubeSeq Supreme service.

Sample submission:

• Order our LightRun 96 barcodes first and label your 96 well PCR plate with V-shaped wells with the respective barcode label
• Use our LightRun 384 Echo Plate for submitting your samples in 384 well format
• Take 5 µl purified template DNA with either of the concentrations given in below table
• Add 5 µl of primer with a concentration of 5 pmol/µl
• Ensure that the total volume of your premixed sample is 10 µl
• For the LightRun 48, please fill the columns from A1 to H6
• Seal your 96well plates using 8-cap strips to prevent material loss
• Use our adhesive plate foil to seal the LightRun 384 Echo Plate
• Ship samples at ambient temperature to our sequencing lab

Conditions:

DNA templates must be purified. Sequencing reactions cannot be repeated and all reactions will be charged. Samples are not stored. For samples containing difficult-to-read streches (GC-rich, hairpin structurs...) we recommend our PlateSeq Supreme service.

 
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
• Label your enclosed primers with our free Tube Labels

Sample storage:

Premixed samples are not getting stored.

Data storage:

Sequencing data are provided in the following file formats by default and are available online for 6 months:

• .ab1 files - ABI trace files with all available raw data
• .scf files -Trace files with base calls and quality values
• .seq files - Clipped & unclipped FASTA sequences
• .phd.1 files - Contains base call and quality information
• .pdf quality report - Trace file with quality scores
• FASTA file with all sequences