Cells were centrifuged, washed and dissolved in genomic DNA isolation buffer (0.4 M NaCl, 0.2% SDS, 0.1 M Tris pH 8.3, 5 mM EDTA).
DNA was dissolved in doubly distilled H20, DNA concentration was estimated by measurement of OD260, and purity was measured by the ratio of OD260/280.
For control injection (mock injection), a solution containing 40 μl of FuGENE™6 dissolved in PBS (without DNA) or PBS only was prepared.
DNA pellets were dissolved in 100 μl of DNA hydration solution.
First, DNA was dissolved in ultrapure water, and Ag-DNA was prepared by mixing 42 ng/μl DNA with 0.0005, 0.015, 0.15, 0.3, 0.75, 1.5, 15 mM AgNO3 according to 1 1 proportion for 15 min, respectively.
DNA was dissolved in 40 µl of Puregene DNA Hydration Solution (Gentra Systems, Valencia, CA, USA).
Genomic DNA was then dried and dissolved in 20 µl of DNA hydration solution.
DNA was then reprecipitated and dissolved in deionized water, and DNA concentration was measured using Eppendorf Biophotometer (Germany).
The pre-hybridization mixture consisted of 400 μg single stranded sheared herring sperm DNA and 125 μg C0t-1 dissolvedlved in 150 μl hybridization buffer.
"DNA normally stays dissolved in water, but when salty DNA comes in contact with alcohol it becomes undissolved," explains the University of Utah Genetic Science Learning Center in a similar tutorial.
