Exosome Purification, Exosomal RNA Purification, and Small RNA-Sequencing Services

Requirements:

BACKGROUND

With collaboration with the National Institute on Alcohol Abuse and Alcoholism (NIAAA), The National Institutes of Health (NIH), Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), requires laboratory services in support of a clinical research study (POTS) in the laboratory of Dr. Paule V. Joseph at the NIH (Building 10, Bethesda, Maryland). The study requires processing of plasma specimens from 31 enrolled human research participants to isolate intact exosomes, purify exosomal RNA, and perform small RNA-sequencing for downstream analysis. These time-sensitive, irreplaceable specimens must be professionally processed to preserve sample integrity and yield analysis-ready data. The requiring laboratory does not have the in-house validated workflow or capacity to perform integrated exosome isolation, exosomal RNA purification, and Illumina small RNA-sequencing to the required standard within the necessary timeframe. The acquisition is therefore pursued to obtain these specialized services from a qualified provider so that the study's analyses can proceed without loss of specimen integrity or previously collected clinical data.

SCOPE OF WORK/TASKS

The contractor shall furnish all labor, equipment, instrumentation, reagents, and materials necessary to isolate intact exosomes, purify exosomal RNA, and perform small RNA-sequencing on the 31 Government-furnished plasma specimens, maintaining specimen integrity and chain of custody throughout.

The following tasks are required for this requirement:

• Purify intact exosomes from 0.5 mL of plasma for each of 31 samples; split the elution into aliquots A and B, and further split aliquot A into two intact-exosome aliquots (two vials per sample returned to the client).

• Purify exosomal RNA from intact exosomes (aliquot B); elute into two RNA aliquots per sample (one returned to the client and one used for small RNA-sequencing); perform QC of the purified RNA.

• Perform small RNA-sequencing for 31 samples, including library preparation, library QC, and sequencing on the Illumina platform (1x51 bp, approximately 20M average raw reads per sample).

• Perform basic bioinformatics analysis (trimming, filtering, and mapping) and transfer all sequencing data to the client via hard drive and digital download.