Center for Clinical and Translational Metagenomics, Request form, v 1.9 03/23/2015 Services Request Form: Center for Clinical and Translational Metagenomics Please fill out relevant sections per experimental work to be undertaken. You may indicate “pending” for IRB/IACUC/COMS protocols, as well as grant and charge-back information. Core staff use this information to identify resources needed for projects, set timelines, and generate quotes for services. Staff of the core will be in contact to discuss specifics and help define best approaches. Prior to the start of a project the investigator must provide: (1) copy of IACUC or IRB protocol, (2) protocol approval letter and (3) Peoplesoft# or PO#. Services requested: please check all areas that apply: Microbiology Gnotobiotic Mouse Facility Molecular and Sequencing Services Computational Services A: Investigator Information: Date of submission: ____________ Investigator (PI): __________________Phone: _____________ Email: ____________ Primary Contact: _______________ Phone: _____________ Email: ____________ Mailing address ______________________________________________________ Billing address _______________________________________________________ Institutional Affiliation: _______________________ Funding Source: _________________ HDDC member: Yes No Grant#:_____________________________ Peoplesoft# (Partners investigators)__________________________________________ Purchase Order# (for non-Partners investigators, can be provided after quote is issued)_______________________________ Protocol information (if submitting human samples, requesting mice or isolates): Human Research Studies: IRB Protocol# ______________________ Expiration Date: _____________________ IRB Granting Institution: __________________________________________________ Animal Studies: IACUC Protocol#: _____________ Expiration Date: ___________________________ IACUC Granting Institution: _______________________________________________ COMS/IBC Protocol#: ___________________ Expiration Date: __________________ (COMS = Committee on Microbiological Safety) COMS Granting Institution: _______________________________________________ Email to vyeliseyev@partners.org or FAX to 617-264-6898, ATTN: CCTM Page: 1 © 2014, Center for Clinical and Translational Metagenomics, Brigham & Women’s Hospital. All rights reserved. Center for Clinical and Translational Metagenomics, Request form, v 1.9 03/23/2015 B.: Description of experiment (add additional pages as needed). Please include (1) aims of experiment (2) source of materials to be submitted for analysis, (3) numbers of biological replicates in each group, (4) desired analyses, (5) how data will be used, and (6) any deadlines. Email to vyeliseyev@partners.org or FAX to 617-264-6898, ATTN: CCTM Page: 2 © 2014, Center for Clinical and Translational Metagenomics, Brigham & Women’s Hospital. All rights reserved. Center for Clinical and Translational Metagenomics, Request form, v 1.9 03/23/2015 C. Gnotobiotic services (1)Mouse strains to use: BALB/c C57BL/6 Swiss-Webster Other: _____________________________________________________________ (2)Mouse gender and age required for the experiment: (3)Source of mice: CCTM bred Taconic Other: _____________________________________________________________ Biohazard level: Procedures: Gnotobiotic BL-1 BL-2 Colonization/gavage Blood collection Gavage of non-microbial materials IP injections Fecal pellet collection Weighing mice in isolators Altered Diet (state desired diet(s): _______________________________________ ___________________________________________________________________ Release GF mice to another facility (list the facility and contact): _______________ _____________________________________________________________________ Other: _____________________________________________________________ D. Samples to be submitted for microbiology, metagenomic or histopathology Species from which materials will be obtained: human mouse Other:______________________________________ Sample type(s): (check all that apply) Stool Whole blood Stomach Skin Duodenum Microbial Isolate Urine Sputum Jejunum Other:_______________________ Ileum Cecum Colon Other: _____________________________________ Sampling method: Email to vyeliseyev@partners.org or FAX to 617-264-6898, ATTN: CCTM Page: 3 © 2014, Center for Clinical and Translational Metagenomics, Brigham & Women’s Hospital. All rights reserved. Center for Clinical and Translational Metagenomics, Request form, v 1.9 03/23/2015 Fecal Pellet/Stool Biopsy Bowel – full thickness Scraping Swab Other:____________________________________________________________ Environmental samples: ___________________________________________ How many samples will be submitted of each type (provide any additional description regarding different sample sets across experiments or for specific types of analyses): Email to vyeliseyev@partners.org or FAX to 617-264-6898, ATTN: CCTM Page: 4 © 2014, Center for Clinical and Translational Metagenomics, Brigham & Women’s Hospital. All rights reserved. Center for Clinical and Translational Metagenomics, Request form, v 1.9 03/23/2015 E. Analyses: Microbiology Qualitative culture – presence/absence of one or more species: Please indicate species of interest: ________________________________ ____________________________________________________________ Quantitative culture – counts only Quantitative culture – speciation of isolates Indicate if specific species should be isolated: ________________________ _____________________________________________________________ Other microbiological analyses: _________________________________ ____________________________________________________________ Do you need stocks of isolates to be prepared from cultured materials? Metabolic profiling Short chain fatty acid analyses Indicate sample type(s) for analysis Pure isolates Stool samples (human or mouse) Cecal contents (mouse) Other: _______________________ Biochemical typing of pure isolates Indicate type of analysis desired for cultured isolates: __________________ _____________________________________________________________ Resistance testing of pure isolates Indicate desired antibiotics to test: _________________________________ ____________________________________________________________ Email to vyeliseyev@partners.org or FAX to 617-264-6898, ATTN: CCTM Page: 5 © 2014, Center for Clinical and Translational Metagenomics, Brigham & Women’s Hospital. All rights reserved. Center for Clinical and Translational Metagenomics, Request form, v 1.9 03/23/2015 Metagenomic and Genomic Analyses Depending upon desired analyses, sample preparation may be undertaken in the CCTM core lab or by the sequencing core providing services. Contact CCTM staff if you have questions about options. DNA extraction and QC RNA extraction and QC 16S rRNA gene library prep for phylotyping and community structure analyses. full-length 16S or 18/28S rRNA gene sequencing for typing (pure bacterial or fungal isolates only). Genome sequencing of pure isolates. Please indicate any preferences for sequencing platform: Illumina MiSeq (250bp paired-end; 16S V4 region, or limited metagenomic whole shotgun reads from DNA or RNA) Illumina HiSeq (Commonly 100-150bp paired-end; metagenomic WGS, RNAseq) Other: ______________________________________________ Core facility for sequencing: CCTM: MiSeq analyses Other: ______________________________________________ Desired read depth/sample: requested average number of reads/sample. You may speak with CCTM staff if uncertain regarding required depth for particular applications. Avg. reads/sample: _____________________ Email to vyeliseyev@partners.org or FAX to 617-264-6898, ATTN: CCTM Page: 6 © 2014, Center for Clinical and Translational Metagenomics, Brigham & Women’s Hospital. All rights reserved. Center for Clinical and Translational Metagenomics, Request form, v 1.9 03/23/2015 Computational Analyses: Discussion with core staff is needed to identify the best approaches for analyzing your metagenomics data. In addition to the general categories below, please indicate desired use of data obtained from analyses. Please briefly describe the data set to be collected and specific hypotheses to be tested. If this is an exploratory project, please briefly describe what sorts of structure you expect to find in the data. Bioinformatics Analyses: 16S rRNA phylotyping - clustering to operational taxonomic units (OTU) with taxonomic assignment, and number of counts/OTU. Whole genome shotgun sequencing of complex communities - assignments of reads to reference genomes or gene families/homologies Genome sequencing of pure isolates: assembly of short reads into contigs, basic genome annotation. Other: _____________________________________________________ Statistical and Machine Learning Analyses: Experimental design and power analysis Prediction of host phenotype from microbial community structure Testing differences among subgroups based on microbial community structure Integration of multiple data sources (e.g., immunological readouts with microbiology results) Relational/network analyses Time-series analyses Other: _____________________________________________________ Email to vyeliseyev@partners.org or FAX to 617-264-6898, ATTN: CCTM Page: 7 © 2014, Center for Clinical and Translational Metagenomics, Brigham & Women’s Hospital. All rights reserved.