10X RBC Lysis Buffer is used for the differential lysis of red blood cells (RBC) present in whole blood.

Applications

BioPioneer’s RBC Lysis Buffer is versatile and suitable for a range of applications:

• 10X RBC Lysis Buffer is used for isolation of DNA and RNA from blood cells.

Composition

The RBC Lysis Buffer is made from high-purity Ammonium chloride, Sodium Bicarbonate, EDTA.

Preparation Instructions:

Preparation of sample (For Erythrocyte Lysis): Mix 1 volume of whole blood with 5 volumes of 1X RBC Lysis Buffer Solution (R075) in Micro Centrifu ge Tube-B (1.5 ml) (provided).For example, to 200 μl of whole blood, add 1 ml of 1X RBC Lysis Buffer Solution.

For optimal results the volume of mixture (Blood + 1X RBC Lysis Buffer Solution) should not exceed ¾ of the volume of the tube to allow efficient mixing. For example, add 5 ml of 1X RBC Lysis Buffer Solution to 1 ml of whole blood, and mix in a tube which has the total volume of ≥8 ml. Whole Blood treated with any common anticoagulant such as heparin or EDTA can be used in this protocol.

NOTE: Use an appropriate amount of whole blood. Upto 1.5 ml of healthy blood (typically 4000‐7000 leukocytes per microliter) can be processed. Reduce amount appropriately if blood with elevated numbers of leukocytes is used.

Incubate for 10‐15 minutes on ice. Mix by vortexing briefly 2 times during incubation. The cloudy suspension becomes translucent during incubation, indicating lysis of erythrocytes. If necessary, incubation time can be extended to 20 minutes.

Centrifuge at 400 x g (1700 rpm) for 10 minutes at 4°C, and completely remove and discard supernatant.

NOTE: Leukocytes will form a pellet after centrifugation. Ensure supernatant is completely removed.

Repeat lysis step with the cell pellet, by adding 2 volumes of 1X RBC Lysis Buffer Solution per 1 volume of whole blood used in step 1. For example, to 200 μl of whole blood, add 400 μl of 1X RBC Lysis Buffer. Solution. Thoroughly vortex to resuspend the cells.

Centrifuge at 400 x g (1700 rpm) for 10 minutes at 4°C, and completely remove and discard supernatant.

Resuspend the pellet in the appropriate buffer and proceed with further procedures.

Safety Information

The RBC Lysis Buffer is intended solely for laboratory applications and should not be used for pharmaceuticals, household purposes, or other non-laboratory uses. Always observe proper laboratory safety protocols, including wearing gloves and safety goggles when handling the buffer. It is not compatible with bleach-containing disinfectants.