RNAseq Statement of Work.docx

RNA LIBRARY CONSTRUCTION AND RNA-SEQ of 72 Samples

Statement of Work: The USDA ARS NCCCWA requires RNA library construction, Quality Control and RNA-seq for 72 RNA samples utilizing paired-end, 150 base pair sequencing via Illumina HiSeq 4000 or equivalent in accordance with the following:

1.) Construction of RNA library with polyA selection for each total RNA sample submitted. Verification of library quality for each of the libraries and resynthesis of any substandard library(s).

2.) Each library sample must correspond to an RNA sample and be barcoded such that sequences from individual samples can be uniquely identified. Libraries will be pooled in groups of 12 for sequencing.

3.) Sequencing utilizes a minimum of 6 lanes on the Illumina HiSeq 4000 (or equivalent) platform yielding a minimum of >35,000,000 reads per library.

4.) All 72 samples will be provided in the same shipment. Groups include:

a. Liver: lane 1 = 6 Fed WT, 6 Fed Mutant; lane 2= 6 Fasted WT, 6 Fasted Mutant (24 samples)

b. Muscle: lane 1 = 6 Fed WT, 6 Fed Mutant; lane 2= 6 Fasted WT, 6 Fasted Mutant (24 samples)

c. Fat: lane 1 = 6 Fed WT, 6 Fed Mutant; lane 2= 6 Fasted WT, 6 Fasted Mutant (24 samples)

* All sample groups must be analyzed using the same sequencing platform.

Deliverable for each sample group will be due not later than 60 days from receipt of samples.

Contractor shall provide the following: All sequencing from each group of 24 samples described above shall be provided by Illumina HiSeq 4000 (or equivalent) platform utilizing 150 base pair, paired-end reads. Each sample group must be analyzed using the same sequencing platform.

1. Barcoding information including barcode sequences corresponding to sample names provided.

2. Data quality must be the Illumina specification or equal that >75% of the nucleotides are greater than Q30 for 150bp sequencing.

3. Deliverables: Data will be provided in FASTQ format for each group of samples and be made available on a secure website or sent on external hard drives to USDA-ARSNCCCWA, 11861 Leetown Road, Kearneysville, WV 25430 within 60 days of receipt the samples.