RNA Sequencing & Expression

Tru-Seq Standard RNA-Sequencing: High Coverage (50M pairs)

Standard RNA Sequencing includes sample receipt, plating, poly-A selection and cDNA synthesis, library preparation, sequencing (76bp paired reads) and sample identification QC check (when Sample Qualification of a matching DNA sample is chosen). The product provides library construction using a non-strand specific Illumina TruSeq Protocol and sequence coverage to 50M Paired reads or 50M Total reads (+/- 5%) (depending on the reference sequence supplied). Processing times vary and depend on current demand.

Deliverable
Data delivery will include a de-multiplexed, aggregated Picard BAM file, which will be accessed via the BASS file server system or FTP for non-Broad users..

Input Requirements

  • Funding and compliance requirements must be in place. This includes a valid IRB or letter of non-engagement where needed.
  • Minimum Sample data including Collaborator Participant ID, Collaborator Sample ID, and Gender.
  • RNA with a RIN (or equivalent QC metric) score below 7 will be attempted on risk.
  • RNA, tissue, blood, stool, or cell pellets that preferably yield >250ng of RNA (note extra cost will be applied for extractions). Samples below 250ng (minimum concentration 5ng/uL) and FFPE samples will be accepted at risk, but success is expected with 100ng (minimum concentration 2ng/uL) or greater.

 

Tru-Seq Strand Specific RNA-Sequencing: Low Coverage (15M pairs)

Tru-Seq Strand Specific RNA Sequencing includes sample receipt plating, poly-A selection and cDNA synthesis, library preparation, sequencing (76bp paired reads) and sample identification QC check (when Sample Qualification of a matching DNA sample is chosen). The product provides library construction using a strand specific Illumina TruSeq Protocol and sequence coverage to 15M Paired reads or 15M Total reads (+/- 5%) (depending on the reference sequence supplied). Processing times vary and depend on current demand.

Deliverable
Data delivery will include a de-multiplexed, aggregated Picard BAM file, which will be accessed via the BASS file server system or FTP for non-Broad users.

Input Requirements

  • Funding and compliance requirements must be in place - this includes a valid IRB or letter of non-engagement where needed.
  • Minimum Sample data including - Collaborator Participant ID, Collaborator Sample ID, Gender. 
  • RNA with a RIN (or equivalent QC metric) score below 7 will be attempted on risk.
  • RNA, tissue, blood, stool, or cell pellets that preferably yield > 250ng of RNA (note extra cost will be applied for extractions). Samples below 250ng (minimum concentration 5ng/uL) and FFPE samples will be accepted at risk, but success is expected with 100ng (minimum concentration 2ng/uL) or greater.


Tru-Seq Strand Specific RNA-Sequencing: High Coverage (50M pairs)

Tru-Seq Strand Specific RNA Sequencing includes sample receipt, plating, poly-A selection and cDNA synthesis, library preparation, sequencing (76bp paired reads) and sample identification QC check (when Sample Qualification of a matching DNA sample is chosen). The product provides library construction using a strand specific Illumina TruSeq Protocol and sequence coverage to 50M Paired reads or 50M Total reads (+/- 5%) (depending on the reference sequence supplied). Processing times vary and depend on current demand.

Deliverable
Data delivery will include a de-multiplexed, aggregated Picard BAM file, which will be accessed via the BASS file server system or FTP for non-Broad users.

Input Requirements

  • Funding and compliance requirements must be in place - this includes a valid IRB or letter of non-engagement where needed.
  • Minimum Sample data including - Collaborator Participant ID, Collaborator Sample ID, Gender.
  • RNA with a RIN (or equivalent QC metric) score below 7 will be attempted on risk.
  • RNA, tissue, blood, stool, or cell pellets that preferably yield > 250ng of RNA (note extra cost will be applied for extractions). Samples below 250ng (minimum concentration 5ng/uL) and FFPE samples will be accepted at risk, but success is expected with 100ng (minimum concentration 2ng/uL) or greater.