SUBMISSION OF SAMPLE(S) Date : Manager of the platform: Thierry Jouenne : +33 2-35-14-66-80 Pascal Cosette : +33 2-35-14-63-97 / Laurent Coquet : +33 2-35-14-60-12 David Vaudry : +33 2-35-14-67-60 / Philippe Chan: +332-35-14-67-35 YOUR COORDINATES NAME: FIRSTNAME: LABORATORY: PROFESSIONAL ADRESS : : E-mail : SAMPLE(S) HOW MANY SAMPLES : WHICH KIND OF ORGANISM ARE THE SAMPLES EXTRACTED FROM? (ex. : Bacteria, Plant, Human) : HOW MUCH BIOLOGICAL MATERIAL (EX. : PROTÉIN) AND YOUR METHOD FOR ESTIMATING IT PACKAGING OF THE: SAMPLES *  0,2µL unskirted sheet with 96 pits (PCR ThermoFast ABgene Low Profile sheet)  1,5mL « Eppendorfs » tubes with the annotated reference of each sample on the tube and it cap.  Others : * For a trypsin digestion put the samples into a 96 pits sheet or into 1,5 mL Eppendorf tubes. Version du 20Jan2012 Page 1 sur 4 KIND OF SAMPLE SENT :  Liquid  Lyophilized  Band(s) SDS- PAGE gel  PVDF Membrane  2D gel spot(s)  Others: REQUIRED ANALYSIS:  1D gel (s) (1)  2D gel(s) (2)  Silver nitrate staining (3)  Blue colloidal staining (4)  Picking spot(s) on gel (5)  Trypsic Digestion (6)  Maldi-Tof mass spectrometry (7)  LC-MS-MS sequencing (8):  Nano-LC- ESI-Trap  Nano-LC- ESI-Q-TOF  Nt microsequencing (9)  Others: INCLUDED PICTURE (JPEG or TIF FORMAT) WITH THE REQUEST OF THE GEL :  yes  no ANALYSIS REQUEST AND INTEGRATED GELS TO PrIMS* :  yes  no *Concern the protein identification analysis by mass spectrometry. After creation of your account, we give you a login and a password in order to integrate your electrophoresis gel picture(s) and to annotate the interest spot(s). Then, you can consult the identification results (which are saved on a protected internet server) from any computer and you can also follow the analysis way in real time. WOULD YOU RECOVER YOURS SAMPLES AFTER THE ANALYSYS (the peptidic samples are conserved by default during two years after the digestion).  yes  no  I certify that I have had knowledge of the submission conditions, which are described on the page number 3 of this document, and the pricing of the service. I wish that my analysis request would be realised as part of:  a provision of service (validation after the signed estimate back and the order form)  a collaboration (I commit to quote during scientific communications (ex. : posters, publications) the scientific responsible of the PFP as well as each member of the PFP which has been in charge of the sample’s analysis and the data treatment).  an internal research (only for the members of our laboratory) Date: Date : Version du 20Jan2012 Signatures of the requester and of his manager : Approbation du responsable de la PFP Page 2 sur 4 Submission conditions 1) The registered quantity of proteins and the used kind of acrylamid gel (size, acrylamid %, linear or with gradient) will have to be establishing with the requester according to the sample and the wished next coming analysis. The service includes the preparation of the proteic samples, the acrylamid gel and the SDS PAGE migration. (2) We have a large range of strips showing different gradient of unmoved pH. The pH gradient choice, the registered quantity of proteins, the 1D and 2D migration system will have to be establishing with the requester according to the sample and the wished next coming analysis. The service includes the preparation of the proteic samples in the IEF buffer, the rehydration of the strips, the iso-electric focus and the SDS PAGE. (3)The silver nitrate staining usually realised in our laboratory uses the paraformaldehyd (Identification by LC -MS-MS is yet possible). The service includes the staining and the scan of the gel. The bio-informatics analyses of the gel are not included but they may be done on request (take contact with one of the technical platform responsible to give more information). (4) The service includes the blue colloidal staining and the scan of the gel. The bio-informatic analyses of the gel are not included but they may be done on request (take contact with one of the technical platform responsible to give more information). (5) According to the gels (SDS-PAGE or 2D) and the picking spots number, the cut is realised manually or automatically through a robot (ProXciser from Perkin Elmer). (6) The trypsic digestion is automatically realised through a robot (Multiprobe from Perkin Elmer). In spite of restricting contaminations, spots or gel fraction (2x2mm) could be registered into a 96 pits sheet with conical bottom (PCR ThermoFast ABgene Low Profile sheet) or into 1,5mL “Eppendorf” tubes. (7) The analysis includes the sample’s takeover, the register on Maldi sheet with the matrix and the sample’s analysis by Maldi –Tof spectrometry mass (PrOTOF). Specters and peak lists are supplied to the customer under respectely rtf and txt format. The raw data can be possibly supplied on request. The identification of proteins in bio-informatic research in data bank are payable by the requester. In the case of a service of identification, this one is realised from the Mascot Daemon search engine (version 2.1.3) and the report of identification « Peptide Summary report » is sent under html format. The treatment of the samples doesn’t include the desalination. (8) The platform in proteomics has three LC-MS-MS machine: the nano-LC/Nanospray QTRAP (Applied Biosystems), the nanoLC-chip-Q-TOF 6520 (Agilent) and the nanoLC-chip-ion trap 6340 (Agilent). The protein identification are preferably realised with the nanoLC-chip-ion-trap 6340 (Agilent). The data are treated with the DataAnalysis software (version 3.4, Bruker Daltonic).The identifications of proteins by bio-informatic research in the data bank are included into the service. They are realised through the Mascot Daemon software (version 2.1.3). All the LC-MS-MS data and the identification report « Peptide Summary report » from Mascot under html format are supplied to the requester. (9) A 20pmoles minimum quantity of proteins or peptides is necessary. The samples can be treated under a soluble form or fixed on a PVDF membrane. For the sequencing of proteins or peptides blocked in Nt or insufficient quantity, the support and so the first cycles will be invoiced. Results (amino acid sequence) are transmitted to the customer with the chromatogram under PDF format. Pricing The pricing of the services are available on the internet site of the platform in proteomics (http://plateformeproteomique.crihan.fr/ptp/prestations_tarifs/). An estimate is established before any service between the customer and the PFP. Processing time For information, the average delays of treatment in 2010/2011 after reception of samples were of 15 days for 1D or 2D electrophoresis, the staining, scanning of gels and spot picking, of 9 days for trypsin digestion, of 11 days for Maldi-Tof analyses, of 9 days for LC-MS-MS analyses (ion Trap or Q-TOF) and of 5 days for Nt microsequencing. For the projects using the various activities of the plateform, the delays will be defined between the customer, the responsible of the plateform and the experimenter (who likely will take care of analyses), according to nature and number of samples and according the project; every research project being subjected to delays by financer organisms. Delays established with the customer will however be able to be modulated according to obtained results. Version du 20Jan2012 Page 3 sur 4 Windows reserved to the platform facility Date de réception des échantillons : par : Analyse(s) prise(s) en charge par : Lieu de stockage avant analyse : Processus utilisé(s)  Electrophorèse  ProXPress  Découpe sur gel  Multiprobe  Maldi-PrOTOF  Nom du fichier informatique Expérimentateur - -  Nano-LC-ESI-Q-TOF LC-MS/MS  Nano-LC-ESI-Trap  P492  Séquenceur(s)  P494 *  Absence d’anomalie : résultats obtenus dans des conditions optimales  Anomalie sans conséquence sur les résultats à savoir :  Anomalie avec des conséquences sur les résultats à savoir : Diffusion des résultats : Version papier donné au demandeur le ….. / ….. / ….. Version papier envoyé par courrier au demandeur le ….. / ….. / ….. Transmis par téléphone au demandeur le ….. / ….. / ….. Envoyés par e-mail directement au demandeur le ….. / ….. / ….. Support de sauvegarde (ex. CD/DVD-ROM) envoyé au demandeur le ….. / ….. / ….. Support de sauvegarde (ex. CD/DVD-ROM) pris par le demandeur le ….. / ….. / ….. Transmis au co-responsable de la PFP le ….. / ….. / ….. Transmis au responsable de la PFP le ….. / ….. / ….. Résultats intégrés au PrIMS et consultable par le demandeur prévenu le ….. / ….. / ….. Autre(s) : Autres commentaires : Version du 20Jan2012 Page 4 sur 4 Conditions d’analyse *   