Standard Operating Procedure

Subject: Simulated Sickle Cell Normal and Patient Samples

Date:  11 March 2008

Approved By:  Megan Waugh

Simulated Normal Hemoglobin A (N):

1. Weigh out 0.025g of Allura Red AC (F.W. 496.43)
2. Mix Allura Red with 5mL of di water
3. Add 5mL of 50% glycerol to the dye mixture
4. Mix well

Simulated Sickle Cell Hemoglobin

1. Weigh out 0.01g of Cresol Red (F.W. 404.42)
2. Mix Cresol Red with 5mL of di water
3. Add 5 mL of 50% glycerol to the dye mixture
4. Mix well

Simulated Sickle Cell Carrier Hemoglobin

1. Mix 2.5mL of Normal hemoglobin with 2.5mL of Sickle Cell hemoglobin

Normal sample

1. Aliquot 12μL into clear 0.5mL centrifuge tubes, label N

Patient sample

1. Aliquot 12μL of normal, sickle cell, or sickle cell carrier, label P1, P2, P3, etc.
2. Try to make every third one different, ex. P1-normal, P2-sickle cell, P3-carrier, P4-normal, P5-sickle, P6-carrier, etc.