RNA Sequencing

Standard RNA-Sequencing: Low Coverage (15M pairs)

Standard RNA Sequencing includes plating, poly-A selection and cDNA synthesis, library preparation, sequencing, sample identification QC check (when Sample Qualification of a matching DNA sample is chosen), and data storage. The product provides library construction using a non-strand specific Illumina TruSeq Protocol and sequence coverage to 15M paired reads and 30M total reads.

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.

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

Standard RNA Sequencing includes plating, poly-A selection and cDNA synthesis, library preparation, sequencing, sample identification QC check (when Sample Qualification of a matching DNA sample is chosen), and data storage. The product provides library construction using a non-strand specific Illumina TruSeq Protocol and sequence coverage to 50M paired reads and 100M total reads.

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.

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

Strand Specific RNA Sequencing includes plating, poly-A selection and strand specific cDNA synthesis, library preparation, sequencing, sample identification QC check (when Sample Qualification of a matching DNA sample is chosen), and data storage. The product provides library construction using a strand dUTP Protocol and sequence coverage to 15M paired reads and 30M total reads.

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. Data storage of 5 years is also provided.

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 >5ug of RNA (note extra cost will be applied for extractions). Samples below 5ug (minimum concentration 100ng/uL) and FFPE samples will be accepted at risk, but success is expected with 3ug (minimum concentration 60ng/uL) or greater.

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

Strand Specific RNA Sequencing includes plating, poly-A selection and strand specific cDNA synthesis, library preparation, sequencing, sample identification QC check (when Sample Qualification of a matching DNA sample is chosen), and data storage. The product provides library construction using a strand dUTP Protocol and sequence coverage to 50M paired reads and 100M total reads.

Deliverable
Data delivery will include a de-multiplexed, aggregated Picard BAM file, which will be accessed via the BASS file server system.

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 >5ug of RNA (note extra cost will be applied for extractions). Samples below 5ug (minimum concentration 100ng/uL) and FFPE samples will be accepted at risk, but success is expected with 3ug (minimum concentration 60ng/uL) or greater.