Experiment
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User |
Minimize users or establish interuser reproducibility in advance. |
Temporal |
Harvest cells or kill animals at the same time of day. |
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Harvest controls and experimental conditions on the same day. |
Environmental |
If samples are to be collected in batches, minimize the time between batch collections. |
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Use intraanimal, littermate, and cage mate controls whenever possible. |
RNA isolation and library preparation
|
User |
Minimize users or establish interuser reproducibility in advance. |
Temporal |
Perform RNA isolation on the same day, and avoid separate isolations over several days or weeks. |
|
Handle all samples in the same fashion (e.g., for freeze–thaws). |
Environmental |
Isolate RNA batches and prepare libraries using standard precautions to minimize contamination. |
Sequencing run
|
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Temporal |
Sequence controls and experimental conditions on the same run. |
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