The Sequencing and Fragment Analysis Unit provides its clients with the most advanced technologies in the field of DNA sequencing and genotyping and offers technical support and advice for those clients who wish to introduce these techniques in their field of work, as support for their research in genetic studies.
The latest generation of automatic DNA sequencing equipment is available, allowing different approaches to DNA analysis to be developed by highly qualified personnel.
The Sequencing Unit offers the methodology based on the chemical Sanger reaction and the subsequent migration in automatic capillary sequencers.
Functioning of the service
Interested users may contact the Unit’s staff to request information and/or a quote for the service.
In order to request the service, the user must send the completed sequencing request form by e-mail or fax.
Fragment analysis
- Marked PCRs
- Microsatellites
- AFLPs,
- SNPs
The equipment of the service allows the use of labelling with: 6-FAM™, HEX™, VIC®, NED™, PET®, ROX ™ and LIZ ®
DNA sequencing
This unit can determine the nucleotide sequence of samples using BigDye® Terminator v3.1 chemistry, from:
- DNA cloned into vectors
- PCR product
The Unit also offers the possibility to perform direct electrophoresis of samples previously processed by the user.
Analysis of samples less than or not multiples of 96 shall be charged at a higher cost if the 48-hour turnaround time is maintained. In this case, the rates will be maintained if the delivery time can be delayed.
Fragments
The fragments to be analysed in each well must be pre-mixed, otherwise an extra sample preparation fee will be charged.
Sequences
Both samples and primers must be submitted properly labelled in separate Eppedorf® tubes or 96-well plates. The name should be less than 8 characters and no punctuation marks should be used.
In the case of user-purified PCR products, 1.5µl of specific primer at a concentration of 5µM per sample and 10µl of DNA should be submitted. The recommended concentration at which the sample should be submitted can be found in this table:
|
DNA type |
Concentration |
|
Plasmids (3-20 kb) |
100 ng/ µl |
|
Plasmids (>20 kb) and cosmids |
200 ng/ µl |
|
PCR products up to 500 bp |
50 ng/ µl |
*Please consult for other DNA types
If PCR products are not purified this must be specified, this will incur additional cost and delay in the delivery of results. It is important that the user is assured of the presence of a single DNA template.
If the sample is expected to cause sequencing difficulties, it is important to inform the unit’s staff for advice and assessment of the problem.
Samples (tubes or plates), together with the sequencing service request form, must be sent at room temperature in padded envelopes by post or courier (avoiding Friday and eves of public holiday).
Platform for Sequencing and Functional Genomics
- Research Support Building (CACTUS)
- Rúa de Constantino Candeira, 1. Campus Vida , 15782Santiago de Compostela