The most common question asked in Differential Gene Expression (DGE) experimental design meetings at the CI is; "should we do RNA-seq or microarray processing?". It all boils down to what questions you want to answer and how the data will integrate into the bigger experiment. I have described some of the most common questions that are asked or discussed and hopefully this information will be useful in getting you thinking about the direction you want to go.
- Why are people doing RNA-seq?
- Isn't RNA-seq really expensive?
- What about analysis, does it take longer to analyse RNA-seq data?
- Do I need as many replicates?
- How long does it take?
- How many samples can be processed at once?
1. Why are people doing RNA-seq? RNA-seq data allows you to have a greater dynamic range than microarray. RNA-seq is a digital reading (counting number of reads) and microarray is the analogue reading (fluorescents units) this can be useful if you are looking at the extremes of expression. You may if you wish in the future take your prepared library and do a different type of sequencing and analysis to look for splice junctions and other transcriptional changes. It is important to remember that wanting more than DGE needs a completely different experimental design. For microarray processing you are restrained to the design of the array and what species you want to explore, with RNA-seq there is no restraint as long as you have an adequate reference genome/transcriptome to align your data to. There are lots of technical reasons why you would choose one method over the other but I do not think that you can ignore the fact that RNA-seq is the new technology and people are choosing the method as it fashionable and may seem to be more attractive for publications.