XIT™ Genomic DNA Blood Kit Protocol for Isolation of Genomic DNA (for 5ml blood)
Invert the tube to mix and incubate 2‐3 minutes at room temperature.
Centrifuge 2,000xg for 5 minutes then remove supernatant carefully without disturbing the pellet.
Add 10ml of RBC Lysis Buffer to the pellet and mix.
Centrifuge 2,000xg for 5 minutes then remove supernatant.
Repeat steps 4 and 5 if pellet is not white.
Vortex the tube to resuspend the cells in the residual liquid.
Add 4ml of XIT™ Lysis Buffer to the resuspended cells and vortex vigorously to lyse the cells.
Usually no incubation is required; however, if cell clumps are visible after mixing, incubate at 37°C for 5‐10 minutes or until the solution is homogenous.
Add 900µl XIT™ Protein Precipitation Buffer to the sample and mix by inverting the tube 10‐20 times.
Centrifuge at 2,000g for 5 minutes.
Carefully, transfer the supernatant to a new tube.
Add 4ml isopropanol to the supernatant and mix by gently inverting the sample at least 20‐25 times.
Centrifuge at 2,000g for 5 minutes.
Discard the supernatant and use a pipette to carefully remove remaining liquid without disturbing the DNA pellet.
Add 2ml 70% ethanol and invert the tube twice to wash the pellet.
Centrifuge at 2,000g for 3 minutes.
Discard the supernatant and drain the tube on a piece of clean absorbent paper.
Allow to air dry for 15 minutes.
Add 500µl TE buffer to dissolve the DNA.
Rehydrate the genomic DNA by incubating at 55‐65°C for one hour.
Incubate overnight at room temperature to ensure complete genomic DNA hydration.
Store DNA at 4°C, for long term storage store at ‐20 or ‐80°C.
Add 5ml whole blood to a 15ml centrifuge tube containing 5ml RBC Lysis Buffer.
