The Sanger DNA sequencing laboratory is part of the structural and functional genomics section of INGEMM (group 29 of IdiPAZ) and includes technicians and advanced DNA dideoxynucleotide sequencing equipment (Sanger method) and DNA marker genotyping.
The sequencing and genotyping quality is ensured by annual participation in various quality control exercises and the accreditation of results.
The objective of the Sequencing Laboratory is to provide support to the Institute’s research groups and to other organisations and companies by placing the laboratory’s instrumentation at their disposal. The laboratory also offers technical advising for nonexpert users who wish to introduce these techniques to their work area.
The available equipment is calibrated to detect various polymorphic markers (MSI, QF-CPA, Profi ler, etc.). These calibrations allow up to 5 fl uorescent markers to be used simultaneously
(with one reserved for molecular size). Thus, various allele sizes can be detected in the same sample.
In the sequencing laboratory, requested sequencing reactions are performed using reagents optimised for the dideoxynucleotide chain termination or Sanger method.
The reactions are analysed in automated fl uorescencebased sequencing equipment using capillary electrophoresis (models 3130xl and 3730xl, Applied Biosystems-Life Technologies). This technique is based on the detection of DNA fragments labelled with 4 fl uorochromes corresponding to the 4 types of nucleotides, which emit laser-induced fl uorescence captured by a CCD (Charged Coupled Device) camera connected to a computer that processes the information, which is then visualised by specialised software.
Rocío Mena de la Cruz
Inmaculada Rueda Arenas
María Victoria Gómez del Pozo
Tel: (+34): 912 071 010 (Ext.235/226)
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