I use Chelex100 for extraction and MEGAquick-spinTotal for PCR purification. Some samples were very faint so i need to make salting out. Thanks in advance.
http://www.bio-rad.com/en-cn/product/chelex-100-resin
Dear Sarah Eissa
I believe your answer in my book
https://www.researchgate.net/publication/306106519_DETERMINING_SOME_QTLs_IN_SOME_INDIGENOUS_CHICKEN_BREEDS
Book DETERMINING SOME QTLs IN SOME INDIGENOUS CHICKEN BREEDS
@M. A. El-Sayed, thank you Dr for your reply. I looked into the full text of the paper but can't find the salting out technique.
Dear Sarah Eissa
Blood sampling & DNA Isolation:
A half ml of blood sample was withdrawn from Jugular vein on EDTA tube as anticoagulant (0.2 ml of 0.5 M EDTA). DNA was freshly extracted from whole collected EDTA-blood. Two and half ml of lysis buffer TSTM (20 mM Tris-HCl pH 7.6, 640 mM sucrose, 2% Triton X-100, 10 mM MgCl2) was added to the aliquot. The mixture was centrifuged and the pellet suspended in 150 μl Proteinase K, 1.5 ml nuclei lysis buffer and 110 μl SDS 20%. After overnight incubation at 37◦ C, the proteins were removed by NaCl 6M and the DNA were precipitated by ice cold absolute ethanol. Extracted DNA was diluted with sterilized water in ratio 5:495μl before the optical density (OD),that is, purity and concentration of each DNA was carried out with the use of ultra violet spectrophotometer.
https://www.researchgate.net/publication/277308729_COMPARATVE_CHICKEN_GENOME_ANALYSIS_OF_EGYPTION_LOCAL_BREEDS_AND_DEVELOPED_STRAINS_3-THE_MICROSATELLITE_DISCRIMINATION_BETWEEN_DOKKI-4_GOLDEN_MONTAZAH_AND_SILVER_MONTAZAH_STRAINS
Article COMPARATVE CHICKEN GENOME ANALYSIS OF EGYPTION LOCAL BREEDS ...
Thank you Dr. El-Sayed, but this is blood not saliva. Can this technique be used with saliva?
Thanks in advance.
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