Most of the following DNA extraction protocols are the web appendix for Bush, K.L., Vinsky, M.D., Aldridge, C.L., and Paszkowski, C.A. 2005. A comparison of sample types varying in invasiveness for use in DNA sex determination in an endangered population of Greater Sage-Grouse (Centrocercus urophasianus). Conservation Genetics, 6: 867-870.
I have tested numerous DNA extraction kits and old methods (e.g. phenol chloroform) and have found that nothing comes close to Qiagen for DNA quantity and quality especially when using small amounts of sample. As new protocols are developed, they will be posted on the web site.
Qiagen DNeasy® Tissue Kit - Animal Tissue Protocol Modification
(Qiagen (2003) DNeasy® Tissue Handbook. Protocol for isolation of total DNA from animal tissues, pp. 18-20. QIAGEN (Valencia, California, U.S.A.).
I modified protocols using the Qiagen DNeasy® Tissue Kit (animal tissue protocol) for the following tissue types:
***Success rate in parentheses are for microsatellite loci and the CHD gene (DNA sexing) on polyacrylamide gels***Agarose gels and/or specing your DNA samples DO NOT give you an accurate idea of your DNA quality based on tests that we completed in 2006***
(1) Avian Blood (100% Success Rate)
(2) Fresh Tissue or Autolytic (decomposing) tissue (works for all internal organs and viscera, muscle tissue, toes, and chunks of feet) (100% Success Rate)
(3) Hatched Eggshell Membranes (100% Success Rate)
(4) Predated Eggshell Membranes (100% Success Rate)
(5) Blood/Pin Feathers (100% Success Rate)
(6) Plucked Body Contour Feathers (100% Success Rate)
(7) Molted Body Contour Feathers (99.6% Success Rate - sample size of 3824 molted body, wing, tail, and down feathers)
(8) Plucked Chick Down (100% Success Rate)
(9) Getting the most out of your Qiagen DNA extraction kit
QIAamp® DNA Micro Kit - Isolation of Genomic DNA from Tissue Samples
(Qiagen (2003) QIAamp® DNA Micro Handbook. Protocol for isolation of genomic DNA from tissues, pp. 35-37. QIAGEN (Valencia, California, U.S.A.).
*** SPIN COLUMNS NEED TO BE REFRIDGERATED ***** BUT LEAVE THEM OUT AT ROOM TEMPERATURE FOR 24 HOURS BEFORE YOU PLAN ON EXTRACTING DNA FOR OPTIMAL RESULTS****
We also have modified protocols using the QIAamp® DNA Micro Kit for the following tissue types:
(1) Avian mouth/bucal swabs (100% Success Rate)
(2) Predated Eggshell Membranes (100% Success Rate)
(3) Plucked and Molted Body Contour Feathers (100% Success Rate)
(4) Plucked Chick Down (100% Success Rate)
DNA Extraction Protocols for Museum Specimens/Historic Tissue Samples
(1) Protocols using the Qiagen QIAamp® DNA Micro Kit (100% Success Rate)
