Fecal Cell DNA Extraction || research , Experiment || Isolation of DNA from mouse stool.

hello research students.
I have performed fecal cell DNA isloation experiment.
steps are mentioned here along experimental video. Hope you this video will be helpful to research students.
🦭🤍...........
Protocol: Isolation of DNA from Stool for Pathogen Detection
Before You Start:🫶
✅ Prepare the Following:
• Thermomixer or water bath at 70°C
• Ethanol (96–100%) added to Buffers AW1 and AW2


💪🍃Step-by-Step Procedure
1️Sample Preparation:
• Weigh below 23mg stool sample in a 2 mL microcentrifuge tube.
2️ Lysis & Homogenization:
• Add 1 ml InhibitEX Buffer to all samples.
• Vortex continuously until it mixes well.
3️ Heat Treatment:
• Incubate at 70°C for 5 min (or 95°C for difficult-to-lyse bacteria).
• Vortex for 15 sec to ensure complete lysis.
4️ Centrifugation:
• Spin at full speed (20,000 rcf) for 1 min to pellet stool debris.
• Transfer 200 µl of the clear supernatant to a new 1.5 mL microcentrifuge tube (avoid solid debris).
☻Add first proteinase K in tube then add 200ul clear supernatant.
5️ Protein Digestion:
• Add 15 µl Proteinase K to the supernatant and do pippeting+ botax.
• Add 200 µl Buffer AL, then vortex for 15 sec to mix.
6️ Incubation:
• Heat at 70°C for 10 min to break down proteins.
7️ DNA Precipitation:
• Add 200 µl ethanol (96–100%), then vortex.
8️ DNA Binding:
• Transfer 600 µl of the mixture onto a QIAamp spin column in a 2 ml collection tube.
• Centrifuge at full speed for 1 min and discard the flow-through.
9️ Wash Steps:
• Add 500 µl Buffer AW1 → Centrifuge for 1 min → Discard flow-through.
• Add 500 µl Buffer AW2 → Centrifuge for 3 min → Discard flow-through.
• Final Spin: Centrifuge again for 3 min to remove residual wash buffer.
10 Elution:
• Transfer the spin column to a new 1.5 ml microcentrifuge tube.
• Add 200 µl Buffer ATE directly onto the membrane.
• Incubate for 1 min, then centrifuge for 1 min to elute the DNA.
11 Measured the DNA concentration using NanoDrop spectrophotometer.
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