Fig. 1

The human 3′UTRome cloning pipeline. a We targeted a panel of 1,815 unique human 3′UTRs and successfully cloned and sequence verified 1,461 unique 3′UTRs (80.1 % cloning success). b The forward primers used to amplify 3′UTR genomic loci were anchored within the last exon of each transcript, ending with the gene specific STOP codons. The reverse primers bound 150 nucleotides downstream of the annotated transcript. c Flow chart summarizing the cloning pipeline of the h3′UTRome v1. Genomic PCR was performed using 3′UTR specific primers and the PCR products were shuttled into Gateway® Entry vectors by recombinational cloning. Single cloned colonies were isolated and screened based on the expected 3′UTR length using PCR and gel electrophoresis. Bacterial colonies passing the screen were then re-arrayed, and the cloned 3′UTRs were sequenced using Sanger sequencing method. The sequence verified 3′UTRs were then submitted to the DNASU plasmid repository for public distribution. 3′UTRs that were not successfully cloned were subject to a second pass of cloning. d Electrophoretic analysis of PCR products from the complete h3′UTRome v1. The sizes of 3′UTRs from 1,461 PCR reactions were analyzed on ethidium bromide stained gels