Blood Treatment for Avian DNA Analysis

(from Michael Wink, Germany, 11 Feb 2004).

1. Blood should be drawn into a syringe with EDTA, sodium citrate anticoagulant. Blood should be drawn only by a veterinarian or similarly experienced person. Blood can also be obtained by vein puncture.

2. A minimal amount of blood to be saved would be about 0.1 ml.

3. Evacuate the blood from the syringe into a small screw cap vial (e.g. 1.5 ml Sarstedt microtube or larger tube if more than 0.5 ml is taken).

4. Immediately add an equal volume of blood lysis buffer (0.1 M Tris, 10% EDTA, 2% SDS; pH 7.5). Mix well by shaking. The SDS lyses the cell and nuclear membranes and denatures proteins. The EDTA essentially inactivates nucleases (enzymes that degrade nucleic acids). Alternatively add 1.5 ml 80% ethanol. Make sure that the cap of the vial closes tightly. Label your vial: write name etc on a small piece of paper and wrap it around the vial by cello-tape. Don't write on the vial as this may wear off during transport or in the freezer.

5. The samples should be safe at room temperature, but it does not hurt to keep them frozen or refrigerated when possible.

Tissue Preservation

If an egg fails to hatch, or a nestling dies, or you find a dead bird at the road side, tissue should be preserved:

1. If it is an embryo then place the whole embryo on a petri dish or other clean plate (preferably on ice) and chop into small pieces with scalpel or scissors. If a nestling, or a dead bird liver, kidney, breast muscle, heart or brain would all be worth dissecting out and saving. Also dice finely.

2. Put the diced embryo or tissue into a 1.5 ml or 15 ml tube (depending on the size of the embryo or tissues).

3. Add 10 times of volume of the EDTA buffer or 80-90% ethanol.

4. The samples "should" be safe at room temperature, but, as above, refrigerate or freeze if possible, and transport on ice.

5. It is preferable if tissue or embryos were removed ASAP after death of the egg/chick.

Feather samples
DNA exist in the lower tip of a feather; DNA recovery from feathers often works but not always. If you can obtain blood, this is a much safer method. Feather sampling should be used in cases that other tissue sources are not available

1. Don't touch the shaft with your fingers

2. Store the feather in a plastic bag at 4C or 20 C

3. Alternatively, cut the lower tip of the feather and store it in EDTA buffer