Supreme sequencing in 96well plates
SupremeRun 96 - Cost-efficient high throughput sequencing
SupremeRun 96 is a high-throughput sequencing service for routine Sanger sequencing of 96 samples with the same primer on a plate.
Automated workflows guarantee easy and convenient sample processing. Prepaid SupremeRun 96 barcodes enables transparent sample tracking during the automated sequencing workflows.
- Sanger sequencing of plasmids or PCR fragments in plates
- Reading lengths of up to 1100 Q20 bases
- Easy online ordering with simple invoicing
Please Note:
The SupremeRun Multiprimer sequencing service has been discontinued. We still accept minimum 48 samples in 96well plates with different primers and you can submit these using our PlateSeq Service.
Product details of the SupremeRun 96 service
- High-throughput Sanger sequencing of up to 96 samples on a microtiter plate
- Full 96well microtiter plate sequencing for 96 samples with one primer
- Free use of a wide variety of standard primers
- Sequencing reads automatically processed with superior data analysis software
- Reads with up to 1100 Q20 bases
- Optimised Sanger sequencing protocols for difficult templates (GC-rich sequences, bisulfit samples or low copy plasmids)
- Free storage of DNA and primers
- Repetition of sequencing reactions upon request (free of charge)
Storage of DNA and primer:
- Storage of template DNA for 4 weeks
- Storage of synthesised primers for 4 weeks free of charge
- Storage of synthesised primers for one year at additional cost
- Storage of enclosed primers for 10 days free of charge
Data storage:
- Secure online archive of all selected sequence data for 6 months
Sample preparation & sumbission for the SupremeRun 96 service
- Please use fully skirted PCR plates
- Template concentration must be normalised across the plate
- PCR product size should not vary by more than a factor of 3
- Samples should be sent liquid in a total volume of 15 µl
- Seal your plates using 8-cap strips to prevent material loss
- Stick the SupremeRun 96 plate barcode on the plate edge
- Label your enclosed primer with our free Primer Barcodes
- Ship samples at ambient temperature to our sequencing lab
|
Sample type
|
Product length
|
Sample conc.
|
Sample vol.
|
| Plasmid DNA |
< 30 kbp |
50-100 ng/µl |
15 µl |
| Purified PCR products |
150-300 bp |
1 ng/µl |
15 µl |
| |
300-1000 bp |
5 ng/µl |
15 µl |
| |
1000-3000 bp |
10 ng/µl |
15 µl |
| Unpurified PCR products |
150-300 bp |
min. 4 ng/µl |
15 µl |
| |
300-1000 bp |
min. 10 ng/µl |
15 µl |
| |
1000-3000 bp |
min. 20 ng/µl |
15 µl |
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Please consider optimal primer conditions, concentration and volume as described below.
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
- Primer must have a total volume of 20 µl (double distilled water or 5mM Tris-HCl)
- Concentration of primers with wobble bases must be calculated according to the following formula: nX x ConcPrimer