DNA Extraction Protocol from Animal Tissue Blocks

DNA is an important component of all living cells and mainly exists in the nucleus. DNA extraction from animal tissue blocks is a fundamental process in molecular biology and genetics research. The extraction procedure typically involves breaking down cellular and nuclear membranes to release DNA molecules and then separating the DNA from other cellular components.

• The tissue blocks were thawed, and washed with normal saline to remove the blood stain, about 0.5 g of tissue was clipped, put into a 1.5 ml centrifuge tube, and cut into pieces.
• Add 0.45 ml TES and mix well, then add 50 μl SDS (10%), and 5.0 μl protease K (20 mg/ml), mix well, hold at 56°C for 4-6 h, and shake once every 2 h.
• Place at room temperature, add equal volume saturated phenol (500 μl), reverse mix, 10000 r/min, the centrifuge 10 min, separate water phase and organic phase, and carefully absorb the upper water phase containing nucleic acid, into a new 1.5 ml centrifuge tube.
• Add equal volume phenol: chloroform: isoamyl alcohol (25: 24: 1), mix upside down, 10000 r/min, centrifuge for 10 min, remove the upper layer, and transfer to a new 1.5 ml centrifuge tube.
• Add equal volume chloroform: isoamyl alcohol (24: 1), mix upside down, 10000 r/min, centrifuge for 10 min, and take the supernatant into a new 1.5 ml centrifuge tube.
• Adding 2.5 times the volume of -20°C pre-cooled anhydrous ethanol precipitated DNA and observed the phenomenon.
• 12000 r/min, centrifuge for 10 min, discard ethanol.
• Stored at -20°C, 75% ethanol was washed, 10000 r/min, centrifuged for 5 minutes, ethanol removed, and DNA dried at 55°C.
• Add the proper amount of TE to dissolve DNA (depending on the amount of DNA) and store at -20°C for later use.

Creative Bioarray Relevant Recommendations

• Creative Bioarray provides a range of normal and diseased tissue blocks. Our high-quality tissues are banked under strict collection protocols to ensure that both paraffin-embedded and frozen tissues are all well-characterized, clinically annotated tissues needed for critical experiments.
• We provide a series of nucleic acid extraction kits for a variety of samples to help our customers accelerate their research. Beyond that, we also offer extraction services of nucleic acid from a wide range of starting materials.

• The force at each step of extraction should be gentle to prevent damage to DNA by mechanical shear force.
• When taking the supernatant, be careful not to pick up the protein layer in the middle.
• When the ethanol is rinsed away, do not shake the DNA.
• After centrifugation, do not shake the centrifuge tube, hold the tube to be stable, and the slope faces outward.

Tissues

Nucleic Acid Extraction