Please download here the protocol for sample preparation.
Steps for Sanger sequencing:
1.- Sample preparation:
When the input material is DNA from PCR, we recommend to confirm the presence of a single PCR product by electrophoresis in an agarose gel. Before sequencing, it is mandatory to purify the PCR product. We offer the PCR purification service for 96 sample plates (see pricing list).
When the input material is plasmid DNA, it is not necessary to perform the DNA purification before the sequencing reaction.
2.- Sequencing reaction (download the sequencing reaction protocol).
Reagents: you can get Big Dye Terminator v3.1 and sequencing buffer (5X) at our core facility.
Samples can be processed in eppendorf tubes or in plates. For less than 50 samples iwe recommend using eppendorf tubes of 1.5 ml. Please label them with your user ID and sample name, and use the same code in the service request form. For more than 50 samples we recommend using 96-well plates.
3.- Sample delivery to the Core Facility:
Sequencing applications are filled online (DNA sequencing application form). Please bring a signed copy of the form along with your samples to the core facility. Applications not generated by the online system are not considered.
4.- Sequencing results delivery:
The Genomics Core Facility will contact you by email and sequence files will be transferred to a central server accessible from any computer.
Result files transferred to the server are automatically deleted every Sunday. Please contact us in case you need older files.
There are two methods to download the generated data:
a) Users connected to the UPF net (Novel) can download the files at: P:\PAS\SCT\Genomica\Sequenciacio
b) The rest of users may connect with the web browser to : https://webfiles.upf.edu/NetStorage
Log on as user: .sctweb.ext.cm.ed.upf and password (ask to the Genomics Core Facility)
Open NetStorage; [email protected] ; Genomica i Sequenciacio
Download and save result files to your computer.