Summary

Estimated time: 10 minutes

• A good experimental design is vital for a successful experiment. If you’re planning to work with a data analyst or bioinformatician, include them in the design stage.

• Aim to minimise variability by identifying possible sources of variance in your samples.

• Biological replicates are important. The number of biological replicates you should have should never be below 3. Technical replicates are often unnecessary.

• Pilot studies are highly recommended for identifying how many replicates and how many reads you should have for enough statistical power in your experiment.

• For basic RNA-seq experiments, poly-A enriched, single-ended, unstranded sequencing at depths of 10M to 20M is probably what you want.