## How do you calculate RPKM?

Here’s how you do it for RPKM:

1. Count up the total reads in a sample and divide that number by 1,000,000 – this is our “per million” scaling factor.
2. Divide the read counts by the “per million” scaling factor.
3. Divide the RPM values by the length of the gene, in kilobases.

What is RPKM in RNA-seq?

Therefore, RNA-seq isoform quantification software summarize transcript expression levels either as TPM (transcript per million), RPKM (reads per kilobase of transcript per million reads mapped), or FPKM (fragments per kilobase of transcript per million reads mapped); all three measures account for sequencing depth and …

What does RPKM measure?

Here, we argue that the intended meaning of RPKM is a measure of relative molar RNA concentration (rmc) and show that for each set of transcripts the average rmc is a constant, namely the inverse of the number of transcripts mapped.

### What is the difference between FPKM and RPKM?

The only difference between RPKM and FPKM is that FPKM takes into account that two reads can map to one fragment (and so it doesn’t count this fragment twice). TPM is very similar to RPKM and FPKM. The only difference is the order of operations.

What is HTSeq FPKM UQ?

Description. Fragments Per Kilobase of transcript per Million mapped reads upper quartile (FPKM-UQ) is a RNA-Seq-based expression normalization method. The FPKM-UQ is based on a modified version of the FPKM normalization method.

How do you normalize RPKM?

Here’s how you do it for RPKM: Count up the total reads in a sample and divide that number by 1,000,000 – this is our “per million” scaling factor. Divide the read counts by the “per million” scaling factor. This normalizes for sequencing depth, giving you reads per million (RPM)

#### How is baseMean calculated?

baseMean—The average of the normalized count values, dividing by size factors, taken over all samples.

What is the baseMean value?

baseMean—The average of the normalized count values, dividing by size factors, taken over all samples. log2FoldChange–The effect size estimate. This value indicates how much the gene or transcript’s expression seems to have changed between the comparison and control groups.