GWAS Plasma Preparation Protocol

 

Supplies for Blood Collection:

22g Needle

2ml Eppendorf tubes - C-2171

K2EDTA 0.5M- 15575020

Procedure:

 

  1. Lacerate the end of the tail of the rat by puncturing the tip with a 22g needle in order to collect the blood.

  2. Milk the tail by squeezing in a downward motion toward the tip to increase blood flow.

  3. Collect whole blood into a tube so that the final concentration of K2EDTA is 5mM – with a stock solution of 0.5M, add 5ul per 500µl of blood.

  4. Immediately after collecting blood, invert the blood 2-4 times to mix the blood and anticoagulant.

  5. Spin the whole blood samples for 10 min at 2000 g at RT to pellet the erythrocytes.

  6. Immediately transfer 500 µL aliquots of the plasma in to fresh 2 mL labeled eppendorf tubes using a pipette with appropriate filter tip. Care should also be taken to avoid taking any red blood cells over into the plasma. It is best to err on collecting less than contaminating with red blood cells.

  7. Visualize the quality of the plasma on the hemolysis scale below and mark the color of the plasma according to the scale 0-6 on the GWAS Blood collection sheet.

 

 

 

 

 

8.  Special care should be made to ensure that the identification on each vial is legible. Make sure that the pen you use is        permanent ink and will not wash off upon thawing.

9.   Samples should be stored at -80 °C until analysis and all transportation should be carried out under dry ice.

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