Expression profiling by high throughput sequencing
Summary
We compared proteinase K buffer (PKBuffer) vs TRIzol vs column-based (PureLink RNA Micro kit) RNA isolation on frozen mouse organs methods using the high throughput CryoGrid system integrated with PIXUL sonicator. Mice were treated with intraperiteal injection of LPS (model of endotoxin induced sepsis) or PBS (control). After 12 hours mice were euthanized and brains, hearts, kidneys, and livers were harvested, immediately placed on ice, and then frozen in CryoTrays.
Overall design
For each isolation method (Trizol vs PIXUL PK Buffer vs PureLink RNA column): 1 pair of tissue sample replicates was taken for each type of tissue treatment (LPS VS PBS) and each organ type. Replicates are from different mice but with the same LPS/PBS treatment. For 2 isolation methods with 2 replicates of 2 treatment conditions across 4 types of organs = 48 total tissue samples
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