Cemia Services

Overview

CeMIA SA offers premium Sanger sequencing services for both plasmid and PCR product.

Features

  • 3-5 working days turnaround from sample receipt
  • High quality reads on the ABI3730xl
  • Runs for both tubes and plates
  • Free universal primers

General Considerations

Always take time to purify and quantify all types of template DNA accurately.

We recommend purification by using commercially available PCR spin column kits.

Otherwise we offer template preparation and/or PCR product purification as anadditional service. Just indicate which method you prefer (PCR or GEL) in the DNA Sequencing Request Form. In this case, provide us at least 25 μl of your PCR product.

Determine concentrations by checking OD.

The OD 260/280 ratio should be about 1.8. Values lower than 1.7 and higher than 1.9 indicate contaminants in the sample that interfere with the determination of the concentration and might inhibit the sequencing reaction.

We also recommend ensuring the quality of your DNA by running your sample on an agarose gel.

Ensure complete removal of PCR primers to avoid multiple sequencing.
Please submit purified, quantified and concentration adjusted template DNA samples according to the following concentrations:

Provide in a separate tube 5μL at 5pmol/μL per reaction of the primer(s) to be used for Sanger sequencing (18-22bp, GC content of 50-55%, Tm of 55-60 oC). Primers must not contain phosphorylation or fluorescent dyes.
Alternatively, you can provide premixed samples (a mixture of template and primer) in a 96 well plate format according to the following table:

Universal Primers: CeMIA sequencing service provides a range of Universal Primers at no charge to the customer.

Just indicate which primer you require in the DNA Sequencing Request Form.


Submission

Complete all details in the DNA Sequencing Request Form (concentration, type, purification requests etc) and send it via e-mail or enclose it in the parcel. Lack of any information will compromise quality of sequence readout.

On the submission form, provide all necessary contact and charging details.

Send us via e-mail or upload a jpeg picture of the samples after purification (include MW markers) to allow visualisation of the relevant templates.

Use 1.5 ml safe-lock tubes or 96well plates for your samples.

Do not tape or wrap tubes with parafilm. Safe-lock tubes offer perfect sealing and evaporation protection.

Use a water resistant marker for labeling of template and primer tubes.

Check that labeling is clear and simple.

Provide adequate protection within padded envelope.

Do not tape tubes to paperwork or leave loose inside the envelope.


Results

Results can be sent via e-mail.

Electropherograms can be viewed using Editview (Mac platform) or SeqScanner (PC platform) both of which are freely available.

View our Sanger sequencing guide for troubleshooting or contact us at support@cemia.eu for an expert’s opinion.


Related Links: DNA Sequensing Request Form, List Of Universal Primers , Method description and troubleshooting