DNA extraction from tissue

Protocol for extraction of DNA from tissue or embryos.

Proteinase K digestion

 * 1) Mix DNA extraction buffer
 * 2) *98 &mu;l ReagentB
 * 3) *2 &mu;l ProteinaseK
 * 4) *Mix fresh. 100 &mu;l is enough for a small pea size chunk of tissue or one embryo
 * 5) Place small piece of tissue or embryo into a microfuge tube containing 100 &mu;l of extraction buffer
 * 6) Incubate at 50&deg;C overnight

Phenol/chloroform/isoamyl (PCI)

 * 1) Prepare PCI mix
 * 2) One part tris-saturated phenol to one part 24:1 Chloroform:Isoamyl alcohol
 * 3) Shake thoroughly to make emulsion
 * 4) Add one volume of PCI to extracted sample
 * 5) Shake tubes for 10 seconds
 * 6) Centrifuge at max speed for 5 minutes
 * 7) Remove aqueous phase to a new tube
 * 8) Repeat as needed
 * 9) Add one volume 24:1 chloroform:isoamyl alcohol
 * 10) Shake tubes for 10 seconds
 * 11) Centrifuge at max speed for 5 minutes
 * 12) Remove aqueous phase to a new tube
 * 13) 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 &mu;l 70% EtOH
 * 6) Centrifuge at max speed for 2 minutes
 * 7) Pipette out ethanol
 * 8) Airdry pellet
 * 9) Resuspend pellet in MilliQ water

BioCoder version
Following is the DNA extraction from tissue protocol in BioCoder, 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 BioCoder (see Source code). More information about BioCoder 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