Feb 24, 2025

Public workspaceDry blot spot cards - Processing plasma EDTA from Whole Blood

DOI
dx.doi.org/10.17504/protocols.io.x54v9r524v3e/v1
  • Sara Lucas Del Pozo1,2,
  • Kai-Yin Chau 1,2,
  • Jane Macnaughtan1,2,
  • Giuseppe Uras1,2,
  • Roxana Mezabrovschi1,2
  • 1University College London;
  • 2Aligning Science Across Parkinson's
Sofia Koletsi
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DOI: dx.doi.org/10.17504/protocols.io.x54v9r524v3e/v1
Protocol Citation: Sara Lucas Del Pozo, Kai-Yin Chau , Jane Macnaughtan, Giuseppe Uras, Roxana Mezabrovschi 2025. Dry blot spot cards - Processing plasma EDTA from Whole Blood. protocols.io https://dx.doi.org/10.17504/protocols.io.x54v9r524v3e/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 05, 2025
Last Modified: February 24, 2025
Protocol Integer ID: 119994
Keywords: ASAPCRN
Funders Acknowledgements:
Aligning Science Across Parkinson's
Grant ID: ASAP-000420
Abstract
This protocol details the processing of plasma EDTA from whole blood sample.
Guidelines
The protocol needs prior approval by the users' Institutional Review Board (IRB), Institutional Animal Care and Use Committee (IACUC) or equivalent ethics committee(s) as applicable.
Safety warnings
The protocol needs prior approval by the users' Institutional Review Board (IRB), Institutional Animal Care and Use Committee (IACUC) or equivalent ethics committee(s) as applicable.
Before start
The protocol needs prior approval by the users' Institutional Review Board (IRB), Institutional Animal Care and Use Committee (IACUC) or equivalent ethics committee(s) as applicable.
Blood Spot samples
Blood Spot samples
4h 30m
4h 30m
Gently invert the purple cap tube(s) 180º and back 8-10 times immediately after collection. Allow Duration00:20:00 - Duration00:30:00 to cool down prior to sample processing.

30m
Aliquot 5 drops of Amount75 µL of whole blood (EDTA) onto Whatman 903 filter paper. Set the rest of the whole aside.

Dry blood spots for a minimum of Duration04:00:00 at TemperatureRoom temperature .

4h
Store dried sample cards in sealed plastic bags at Temperature-80 °C with a desiccant.

Procedure
Procedure
20m
20m
The tube with remaining blood following the dried blood spot should be gently inverted 180º and back 8-10 times.

Centrifuge the tube at Centrifigation1200 x g, 4°C, 00:10:00 NO BRAKE.

10m
Centrifigation
After centrifugation there should be a clear separation between the red blood cells at the bottom and the plasma at the top.

Use the pasteur pipette provided or a micropipette to carefully aliquot the EDTA plasma layer into a 15L falcon tube. Approximately Amount3 mL /Amount3000 µL of the plasma from the purple cap tube into the falcon tube.

Note
Do not to disturb the red blood cell layer and set the tube aside. If the sediment is disturbed the centrifugation step must be repeated.

Place the falcon tube in the centrifugation at Centrifigation1800 x g, 4°C, 00:10:00 NO BRAKE.

10m
Centrifigation
From the red blood cell layer, use a micropipette to carefully aliquot Amount500 µL of the red blood cells from the purple cap tube into the 4-5 x 1ml cryovials (1000µl into each tube) labelled as RBC.

Pipetting
Retrieve the falcon tube EDTA plasma layer from the centrifuge. Use a micropipette to carefully aliquot Amount300 µL of the plasma from the falcon tube into the 2-3 x 1ml cryovials (Amount1000 µL into each tube) labelled as EDTA-PLA.

Note
Do not to disturb the pellet at the bottom of the tube. If the sediment is disturbed the centrifugation step must be repeated.

Place plasma samples in Temperature-80 °C .