Question: How should I collect blood from isolating PBMCs for single-cell sequencing?
Answer: There are several options for blood collection tubes:
Vacutainer tubes: Whole blood can be collected using vacutainer tubes. In-house we have only evaluated Sodium Heparin, Sodium Citrate, K2EDTA, and ACD-A anticoagulants, and the single-cell data generated is comparable. Users looking to isolate PBMCs can collect samples in these tubes and perform PBMC isolation using a ficoll gradient and SepMate tube ( follow option 3 of our demonstrated protocol Isolation of Leukocytes, Bone Marrow, and Peripheral Blood Mononuclear Cells for Single Cell RNA Sequencing). Vacutainer tubes are also a good option for users who may want to do multiple assays (i.e., Flow, CytoF, ELISA) from the same blood tube.
CPT tubes: Whole blood from which PBMCs (no granulocytes) are to be isolated may benefit from the use of CPT tubes. CPT tubes with Sodium Heparin and Sodium Citrate anticoagulants were evaluated in-house and generated comparable single-cell data. These tubes already contain a density gradient. After blood collection, tubes are spun to separate the sample into layers, and PBMCs are isolated (follow option 2 of our demonstrated protocol, Isolation of Leukocytes, Bone Marrow and Peripheral Blood Mononuclear Cells for Single Cell RNA Sequencing). CPT tubes are a good option for beginners as the density separation is only a 1-step centrifugation process, and only one tube is needed for both collection and PBMC isolation.
Products: Single Cell Gene Expression, Single Cell Immune Profiling, Fixed RNA Profiling Gene Expression