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Arichard:Protocols/MDA on microarray
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====Lysis==== * Mixing is important for uniform array performance. Avoid touching rubber gasket with pipette tip, however. # In order to lyse bacteria, plate must be freeze thawed. This is only necessary for bacteria, but will not harm eukaryotic samples. ## Cover with cover slip. ## 1 minute on dry ice, 1 min off. Repeat 5x total. # Add 4.5 ul ALS to each array, swirl to mix. Incubate 10 min on ice block. Use this time to finish MDA master mix: ## 2.2 ul MDA buffer. ## 5 ul 200 uM N6. ## 0.8 ul 25 mM dNTP. ## 0.08 ul 50X SYBR Green. ## 1.6 ul phi29 Polymerase (Add last, immediately before use). # Put on ice and add 4.5 ul NS, swirl to mix. # Immediately add 9.68 ul MDA master mix, swirl to mix. # Overlay each array with 60 ul mineral oil. Ensure oil covers array. Bubbles are OK.
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