Thanks Venu, but that part I know. I need to know what value to take to calculate the number of fragments. I have also seen the video in the link provided, but it only explains RPKM and TPM.
how to calculate fpkm?
Hi,
I am working with paired-end reads and I have to now calculate FPKM. My question is, how to calculate FPKM. wc -l mcf7_1.fastq gave me 26723524 reads and wc -l mcf7_2.fastq gave me 26723524 reads as well (I divided the wc -l output by 4), but for FPKM, do I need to take only 26723524 fragments or 53447048?
Thank you.
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It is not possible to calculate FPKM from raw fastq files. You need to align the data to a reference genome. Then you'll get a BAM file. There are many tools out there to calculate FPKM using BAM files.
Here is your guide on how to calculate FPKM.
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An update (6th October 2018):
You should abandon RPKM / FPKM. They are not ideal where cross-sample differential expression analysis is your aim; indeed, they render samples incomparable via differential expression analysis:
Please read this: A comprehensive evaluation of normalization methods for Illumina high-throughput RNA sequencing data analysis
Also, by Harold Pimental: What the FPKM? A review of RNA-Seq expression units