DNA extraction from tissue

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Protocol for extraction of DNA from tissue or embryos.

Proteinase K digestion

1. Mix DNA extraction buffer
   • 98 μl ReagentB
   • 2 μl ProteinaseK
   • Mix fresh. 100 μl is enough for a small pea size chunk of tissue or one embryo
2. Place small piece of tissue or embryo into a microfuge tube containing 100 μl of extraction buffer
3. Incubate at 50°C overnight

Phenol/chloroform/isoamyl (PCI)

1. Prepare PCI mix
   1. One part tris-saturated phenol to one part 24:1 Chloroform:Isoamyl alcohol
   2. Shake thoroughly to make emulsion
2. Add one volume of PCI to extracted sample
3. Shake tubes for 10 seconds
4. Centrifuge at max speed for 5 minutes
5. Remove aqueous phase to a new tube
6. Repeat as needed
7. Add one volume 24:1 chloroform:isoamyl alcohol
8. Shake tubes for 10 seconds
9. Centrifuge at max speed for 5 minutes
10. Remove aqueous phase to a new tube
11. Continue to precipitation

Ethanol precipitation

1. Add 2 volumes 100% EtOH
2. Add 1/10 volume 3M Sodium Acetate pH 5.0
3. Centrifuge at max speed for 10 minutes
4. Decant ethanol
5. Add 150 μl 70% EtOH
6. Centrifuge at max speed for 2 minutes
7. Pipette out ethanol
8. Airdry pellet
9. Resuspend pellet in MilliQ water

BioStream version

Following is the DNA extraction from tissue protocol in BioStream, a high-level programming language for expressing biology protocols. What you see here is the auto-generated text ouput of the protocol that was coded up in BioStream (see Source code). More information about BioStream can be found on my home page. Feel free to mail me your comments/ suggestions.Vaishnavi

Text Output

DNA extraction from tissue protocol

Source Code

DNA extraction from tissue protocol - source code

See also

• Oneill Lab:DNA Extraction
• Mouse tissue lysis for genotyping
• DNA extraction - Salting Out

External links

• DNA from fresh or frozen tissue by Ried lab
• DNA from liver by Bowtell and colleagues - cached version from protocol online
• DNA extraction from tail or tissue - at BioProtocol from unknown contributor
• Protocol Online links to DNA extraction protocols