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The cRNA yields were quite different.
The cRNA yields for these four samples ranged from 130 μg to 145 μg, with a mean of 136 μg.
The cRNA yields from the 0.5 μg samples with our protocol averaged 32 μg (± 13 μg), more than enough for the standard hybridizations.
The cRNA yields from the 0.1 μg samples using the same protocol averaged 10 μg with a range from 5 to 21 μg, so some samples had too little to prepare a hybridization mixture at the same concentration.
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The cRNA yield was measured at 260 nm using NanoDrop 1000A Spectrophotometer.
The cRNA yield was determined by measuring UV absorbtion at 260 nm.
The cRNA yield from these samples were in the range of 25±4.1 with an acceptable 260/280 ratio of 1.9 2.1.
Spectrophotometric analysis was used to determine the cRNA yield as well as the quality of cRNA (optical density 260/280 = 2.0 for each cRNA).
The cRNA yield was measured at using RiboGreen RNA quantitation kit (Invitrogen, CA) and 750ng of cRNA sample was hybridized on a human HT-12 expression beadchip (Illumina, CA) profiling 48,804 transcripts per sample.
Because the cRNA yield from the 1 μg pooled sample test was low, we decided to amplify the smaller samples.
Samples were then purified (Qiagen) with the cRNA yield and quality assessed using the nano-drop and Bioanalyser respectively.
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