Feb 26, 2026
Dried blood spot biobanking protocol
- Taylor P. Johnson1,
- Ariel Saurí1,
- Daniel W. Sirkis1,
- Jennifer S. Yokoyama1,2,3
- 1Fein Memory and Aging Center, Department of Neurology, Weill Institute for Neurosciences, University of California, San Francisco, CA, USA;
- 2Global Brain Health Institute, University of California, San Francisco, CA, USA;
- 3Department of Radiology and Biomedical Imaging, University of California, San Francisco, CA, USA
- Taylor Johnson
Protocol Citation: Taylor P. Johnson, Ariel Saurí, Daniel W. Sirkis, Jennifer S. Yokoyama 2026. Dried blood spot biobanking protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.kxygx8qeov8j/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: February 18, 2026
Last Modified: February 26, 2026
Protocol Integer ID: 243708
Keywords: DBS, dried blood spot, whole blood, Capitainer, B50, UCSF, Yokoyama, dried blood spot biobanking protocol, standard whole blood biobanking method, biobanking of whole blood, anticoagulated whole blood, phlebotomy tube anticoagulant volume, total volume whole blood, other analytes through downstream application, biobanking, accurate quantification of nucleic acid, other analyte, analyte, peripheral blood mononuclear cell, blood spot, nucleic acid, stable consistent 50 ul volume db, protocol supplement, standard phlebotomy, protocol supplements original work, ul volume db, pbmc
Abstract
This protocol presents methods for collection and biobanking of whole blood in the form of a dried blood spot (DBS) from anticoagulated whole blood drawn by standard phlebotomy. Leveraging Capitainer B50 cards, for a shelf stable consistent 50 ul volume DBS, this protocol may be inserted in the beginning of any workflow involving fresh anticoagulated whole blood, such as isolation of peripheral blood mononuclear cells (PBMCs), in which only 75-150 ul may be available to spare. Resulting DBS are held at room temperature until elution for downstream applications enabling a more flexible workflow compared to standard whole blood biobanking methods. Accurate quantification of nucleic acid or other analytes through downstream applications may be achieved by correcting through a dilution factor, considering (i) phlebotomy tube anticoagulant volume and (ii) total volume whole blood drawn. This protocol supplements original work published by Capitainer AB, Uppsala, Sweden.
Materials
- 8.5 ml acid-citrate-dextrose (ACD) or ethylenediaminetetraacetic acid (EDTA) phlebotomy tube (whole blood drawn)
- Capitainer® B50 dried blood spot device, Capitainer AB, Uppsala, Sweden
- P200 wide bore tips
- P200 mechanical pipette
- Tissue culture biosafety cabinet
- 50 ml conical
Protocol materials
70% ethanol
Capitainer B50 sampling card pack
Troubleshooting
Problem
The full lane remains red.
Solution
Possible Causes & Solutions:
1. Pipetting too quickly, leading to inadequate sampling: Pipette slowly and steadily.
2. Excess blood left in the pipette tip, reducing the delivered volume: Ensure the full sample is dispensed from the tip. Pipette slowly and steadily.
3. Bubbles introduced from uneven pipetting: Maintain consistent pressure on the pipette plunger to avoid bubble formation in the sampling channel.
Materials preparation
30m
Obtain whole blood drawn into ACD or EDTA phlebotomy tube.
Clean and sanitize tissue culture hood with 70% ethanol , wipe down with Kimwipe, and UV sterilize.
Allow biospecimen to come to room temperature (00:30:00 post-draw) and briefly spin to remove residual whole blood from the top of tube.
30m
Spray phlebotomy tube and sealed Capitainer B50 sampling card pack into tissue culture hood with 70% ethanol .
Transfer total volume of whole blood into labeled 50 mL conical with 10 mL serological pipette tip to ensure whole blood is uniform prior to sampling.
Note total volume of whole blood.
Proceed with PBMC isolation or other primary phlebotomy tube application using the remaining whole blood volume.
Dried blood spot generation
30m
Note
For standard B50 sampling methods, please see Capitainer AB, Uppsala, Sweden, Capitainer‱ B50 dried blood spot device: Instructions for Use, 2025. Available from: Manufacturer website.
Open theCapitainer B50 sampling card pack in the tissue culture hood (use within 00:30:00 of opening).
Label card with relevant project info prior to sampling on top card lid. Use gentle pressure to maintain integrity of fragile sampling channels.
Fold back the protective tab on the bottom of the card and secure in the slit in back to create declined sampling lanes.
Note
Do not touch sample inlets.
Slowly and smoothly pipette 75 µL of whole blood into one card lane until the sample indicator line has been filled. There should be 5 µL (2-5 µl) whole blood remaining in tip.
Let card sit open at Room temperature for 00:30:00 .
Note
Upon successful sampling, only the control window should be red.
30m
After the 00:30:00 wait period remove the adhesive seal on the bottom tab of theCapitainer B50 sampling card pack and collapse the folds to protect the sample inlets.
Label with any additional study info and tape closed for storage at Room temperature .
Record all observations and storage locations in secure spreadsheet or LIMS database
system
Protocol references
Capitainer. (2025). CAPITAINER-B50 instructions for use [PDF]. Capitainer. https://capitainer.com/wp-content/uploads/2025/07/CAPITAINER-B50_IFU_EN.pdf