DNA sequencing



SUPAMAC has built up a strong reputation in the quality and reliability of its DNA sequencing service. Contact Dr Nicole Sawyer for information about this service and your specific needs.
At SUPAMAC we attempt to meet your needs with state of art analysers such as the ABI 3730 platform
Most sequencing projects are completed within 24-48 hours, and a commercial-in-confidence philosophy is maintained at all times.

Practical Notes
Please read the following notes carefully in order to profit from our sequencing service. Call us on 02 95158403 if you have any questions

  

Information
 
   

Services
Plasmids
Host strain: Please use endA- E. coli strains (XL1-Blue, DH5_, DH1, C600, SURE, NM294). Other strains like JM83, JM101, NM522, NM544, TB1, TG1, BL21, MC1061, Y1088 yield DNA of low quality resulting in sub-optimal sequencing results.
  
 
Plasmid isolation
The success of your sequencing reactions depends on the quality that YOU provide us with. SUPAMAC recommends the Marligen (old Concert) or Viogene kits from Diagnostic Technology (10% and 15% discount on the kits respectively for customers of SUPAMAC). Estimate concentration on an agarose gel, loading a known amount of size marker for comparison. Estimations from a gel are more accurate than OD260 values.
  

Links
 
   

PCR products
To achieve optimal results, you should eliminate the nucleotides and the primers. SUPAMAC recommends the Marligen (old Concert) or Viogene kits from Diagnostic Technology (10% and 15% discount on the kits respectively for customers of SUPAMAC). It is necessary to quantify the purified product by gel electrophoresis, with a size marker of known concentration. Please send us your estimation of the concentration. In some cases, it is a good idea to use internal sequencing primers.


Preparing samples for sequencing
Dye-Terminator chemistry uses labelled ddNTPs to sequence PCR products and plasmids. DNA and primer should be provided as a cocktail.

PCR products
Template Quantity
Primer + H2O
100-200 bp
2-6 ng
5-10 pmol (all samples)
200-500 bp
6-20 ng
& dH2O as required
500-1000 bp
10-30 ng
to a final volume
1000-2000 bp
20-80 ng
of 16ul for all samples
>2000 bp
Plasmid DNA
Template Quantity
Primer + H2O
200 – 500 ng
As above


Please download an order form and email it to support@supamac.com. Put your name/initials and the date in the file name. If you do not email a copy of the order form your samples will not be processed until an order form has been received. Please also include a hard copy of the order form with your samples.
What is the turnaround time?
Samples dropped off at SUPAMAC before 4:00 pm will be processed on the following business day subject to space availability. The results will generally be available on the second business day following sample drop off. For high quality DNA, the guaranteed read length is 700 bases. Sequences are emailed as text files with or without electropherograms (your choice). Hard copies are put in the post.