I used Qiagen Tissue & Blood DNA extraction kit for Red Tilapia (Freshwater fish). I used fin samples. The results were good and proceed with microsatellite analysis.
The Qiagen Promega kit should be okay for you but you must try and search to see if the yield and quality will be the same as for your kind of sample since normally it is used for human blood samples...
Courtesy: Ekrem Ayna · Mie University
FROM FISH BLOOD
Using the blood samples fixed in ethanol, 70μl of each solution was aliquoted into
sterile Eppendorf tubes (Thermo Life Sciences). Each tube was placed in a
centrifuge (MSE Micro Centuar) and pulse spun at maximum speed for 30 seconds.
Excess ethanol was taken off the top of the solution and the tubes were left open to air dry on the bench for 5-10 minutes.
After the ethanol had evaporated, 10μl proteinase K (ABgene, 20 mg/ml) and 340μl
of TEN (50mN Tris HCl pH 8.0; 100mM EDTA pH 8.0; 100mM NaCl; 1% SDS(Sodium Dodecyl Sulfate)) buffer was added to each tube. The SDS causes the cells to rupture and initiates protein denaturation while the proteinase K reduces proteins to their component amino acids. The tubes were vortexed for 30 seconds and incubated at 55oC in a rotating oven (Techne Hybridiser HB-1) for 4-6 hours.
After incubation 10μl RNAase (DNAse Free, Abgene, 2mg/ml) was added to each tube and incubated at 37oC, also in a rotating oven, for one hour in order to digest and remove traces of RNA.Following the incubation period 370μl of phenol was added to each tube to extract the denatured protein from the DNA solution. The solutions were mixed vigorously for 10 seconds and shaken gently every 5 minutes for 20 minutes.
After the 20 minute period an equal volume of chloroform was added to the tubes to absorb and eradicate traces of phenol. The solutions were again mixed vigorously for 10 seconds and then shaken gently every 5 minutes for another 20 minutes. Each tube was then spun for 5 minutes at 10,000g. This causes the solutions to separate into two distinct layers. 270μl of the top aqueous layer, which contains the DNA, was removed and pipetted into a new tube, using a wide-bore tip. 870μl of 92% ethanol (Sigma) was added to the aqueous solutions and each tube was mixed by vigorousinversions, causing the DNA to precipitate out of the solution. After 3 minutes the tubes were pulse spun (10s at 10,000g) to form a DNA pellet at the bottom of the tube.
The ethanol was decanted off and 1ml of 70% ethanol was added to each tube
to wash the pellet. Tubes were placed in a rotator overnight and the alcohol was removed using a micropipette, after which they were then left open to air dry for 10-15 minutes. The pellets were then resuspended in 100μl TE pH 8.0 (10mM Tris, 1mM EDTA) buffer.
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