1. Add 50uL of 10X DNase I Buffer to 450uL lysate in a 2mL DNA Lobind tube.
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1. Add 50uL of 10X DNase I Buffer to 450uL lysate in a **2mL** DNA Lobind tube.
2. Add 1uL of 1U/uL DNASE I and 1uL of 10mg/mL RNAse A and incubate at 37C for 1.5h (without shaking).
3. Add 20uL of 0.5M ultrapure EDTA to inactivate DNAse I and RNAse A (mix gently)
4. Add 1.25uL of 20mg/mL Proteinase K to the mix, mix gently, and incubate at 56C for 1.5h (without shaking).
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5. Add 525uL AL Buffer to the treated ~522 ul treated phage lysate, vortex, and incubate at 70C for 10 minutes.
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6. Add 525 uL 100% EtOH and vortex to mix.
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7. Transfer 750uL of the mix to a DNeasy Mini spin column in a 2mL tube (do it multiple times until all the liquid is through)
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8.
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5. Add 523uL AL Buffer to the treated ~523 ul treated phage lysate, vortex, and incubate at 70C for 10 minutes.
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6. Add 523 uL 100% EtOH and vortex to mix.
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7. Transfer 750uL of the mix to a DNeasy Mini spin column in a 2mL collection tube and centrifuge for 1 minute at 6000x rcf. Discard flow through. Do it multiple times until all the liquid has been passed through the column, discard flow-through and collection tube.
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8. Place the column in a new 2mL collection tube, add 500uL buffer AW1 and centrifuge for 1min at 6000x rcf. Discard flow through with collection tube.
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9. Place the column in a new 2mL collection tube, add 500uL buffer AW2 and centrifuge for 3min at 20,000x rcf to dry the membrane. Discard flow through and collection tube. Centrifuge again with a new collection tube for 1min at 20,000x rcf to make sure there's no carry over buffer left.
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10. Place the DNeasy spin column in a sterile 1.5mL DNA LoBind tube and elute with 30uL AE Buffer or water (heated to 65C, incubate the tube with the water/buffer at 65C for increased yield for 5 minutes).
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11. Spin the column at 6000x rcf for 1 minute to elute DNA
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12. Do a second elution with 30uL of buffer/water to increase yield even further. Store at 4C.
