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. Author manuscript; available in PMC: 2015 Jan 1.
Published in final edited form as: Nat Protoc. 2014 Jun 26;9(7):1740–1759. doi: 10.1038/nprot.2014.121

Figure 1. Detailed experimental workflow of TIF-Seq.

Figure 1

The protocol is composed of: (a) 5′RNA oligo-capping using a DNA/RNA oligo of known sequence (5RNAGsuI), (b) generation and PCR amplification of full-length cDNA, (c) intramolecular circularization after NotI digestion, and (d) purification of paired-end cDNA tags followed by generation of Illumina-compatible sequencing libraries. Individual steps are labeled in black and the structure of the final sequenced library is depicted in the bottom right side of panel d. CIP stands for Calf Intestinal Alkaline Phosphatase, TAP for Tobacco Acid Pyrophosphatase, and ssRNA for single-stranded RNA. Refer to the main text for specific details.