XIT™ Genomic DNA Blood Kit Protocol for Isolation of Genomic DNA ( for 0.5ml blood )
Invert the tube to mix and incubate 2‐3 minutes at room temperature .
Centrifuge 14,000xg for 30 seconds then remove supernatant carefully without disturbing the pellet .
Add 1ml of RBC Lysis Buffer to the pellet and mix .
Centrifuge 14,000xg for 30 seconds 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 400µl 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 90µl XIT™ Protein Precipitation Buffer to the sample and mix by inverting the tube 10‐20 times .
Centrifuge at 16,000g for 5 minutes .
Carefully , transfer the supernatant to a new tube .
Add 400µl isopropanol to the supernatant and mix by gently inverting the sample at least 20‐25 times .
Centrifuge at 14,000rpm for 5 minutes .
Discard the supernatant and use a pipette to carefully remove remaining liquid without disturbing the DNA pellet .
Add 200µl 70 % ethanol and invert the tube twice to wash the pellet .
Centrifuge at 14,000rpm for 5 minutes .
Discard the supernatant and drain the tube on a piece of clean absorbent paper .
Allow to air dry for 15 minutes .
Add 50µl TE buffer to dissolve the DNA .
Rehydrate the genomic DNA by incubating at 55‐65°C for one hour .
Store DNA at 4°C , for long term storage store at ‐20 or ‐80°C .
Incubate overnight at room temperature to ensure complete genomic DNA hydration .
Add 0.5ml whole blood to a 1.5ml tube containing 1ml RBC Lysis Buffer .
