WoodCock Extractions

Quill test Extractions

Three of the test feathers were digested overnight to ensure digestion was successful prior to starting sample extractions. Feather tips were fully digested after overnight digestion. Test feathers were not fully extracted to DNA.

Digestion Protocol

1. Cut quill or tip of feather and place in a 2mL collection tube.

2. Set incubator to 56C.

3. Add 180uL of buffer ATL, 20uL of Proteinase K, and 10uL of 1M DTT.

4. Vortex and place on 56C incubator for a minimum of 12 hours.

5. After removal, vortex and spin down tubes.

1M DTT Solution Protocol

Received a 5g container of powdered or lyophilized DTT.

DTT molecular weight: 154.25 g/mole

1. Weigh out 3.86g of lyophilized DTT and pour into a 50mL conical tube.

2. Add 20mL of ultra pure water to the 50mL conical tube.

3. Stir solution and mix solution until DTT is fully dissolved.

4. Bring volume up to 25mL with ultra pure water and stir/mix again.

5. Store in -20C freezer.

Note: should aliquot into smaller tubes to reduce excessive freeze/thaw cycles (this has not been done yet).