DNA Extraction from fish fins
- Put 1cm2 of fin tissue to 700ml of TNES-Urea Buffer
- Add 3.5 ml of 10mg/ml Proteinase-K
- Digest O/N at 37 C
Wear gloves and work in fume hood when working with Phenol and
Dispose of waste in Hazardous waste container
- Extract two times with an equal volume of Phenol:Chloroform:Isoamyl Alcohol (25:24:1)
- Add 700ml of P:C:I
- rotate or invert for 10 minutes
- Spin at full speed for 10 minutes (Centrifuge in hood)
- remove the top aqueous layer into new tube.
- repeat with an equal volume of P:C:I
- Extract one time with an equal volume of C:I (24:1)
- Add 1/10 volume of 3M NaOAc pH 5.3
- Add 2 volumes 100% EtOH
- Either (1) collect spool of DNA with a pipette and wash in a new tube of 70% EtOH or (2)
spin for 10 min. at full speed, pour off supernatent and wash pellet in 70% EtOH
- Pour or aspirate off EtOH and let pellet air dry.
- Resuspend pellet in water or TE.