XIT™ Genomic DNA Blood Kit Protocol for Purificaton of DNA from Amniotic Fluid
Add 1‐3ml Amniotic fluid to a 1.5ml centrifuge tube.
Centrifuge 14,000xg for 5 seconds then remove supernatant carefully without disturbing the pellet.
Remove supernatant leaving 10‐20µl residual liquid in the tube.
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.
Place the tube on ice for 1 minute to rapidly cool to room temperature.
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 and 5µl Glycogen Solution 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.
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.
