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. Author manuscript; available in PMC: 2016 May 11.
Published in final edited form as: Cold Spring Harb Protoc. 2015 Apr 13;2015(11):951–969. doi: 10.1101/pdb.top084970

Figure 1.

Figure 1

Overview of RNA-Seq. First, RNA is extracted from the biological material of choice (e.g., cells, tissues). Second, subsets of RNA molecules are isolated using a specific protocol, such as the poly-A selection protocol to enrich for polyadenylated transcripts or a ribo-depletion protocol to remove ribosomal RNAs. Next, the RNA is converted to complementary DNA (cDNA) by reverse transcription and sequencing adaptors are ligated to the ends of the cDNA fragments. Following amplification by PCR, the RNA-Seq library is ready for sequencing.