Skip to main content
Figure 3 | BMC Genomics

Figure 3

From: Small RNA pyrosequencing in the protozoan parasite Entamoeba histolytica reveals strain-specific small RNAs that target virulence genes

Figure 3

Small RNA distribution and biochemical analysis of small RNAs for genes with both antisense and sense small RNAs. (A) EHI_020920 (represented by arrow) is depicted with the small RNAs that map to it; each bar represents one unique small RNA (red: small RNAs map to upper strand, sense to EHI_020920; blue: small RNAs map to lower strand, antisense to EHI_020920). The positions of selected probes used for small RNA Northern blot analysis are represented by bars and numbers (black for detecting antisense small RNAs; red for detecting sense small RNAs). The positions of the F and R primers, used to generate cDNA for the strand-specific RT-PCR, are shown. (B) Northern blot analysis detected signal for antisense small RNAs (probes 1, 2, 3) and sense small RNAs (probes 4, 5). (C) Sense small RNAs have 5-polyP termini. 10 μg small RNA enriched sample was treated with Terminator or Capping enzyme. Probes 1, 4, 5 were used for Northern blot analysis. A control oligonucleotide that is labeled with a 5-monoP is degraded by Terminator and has no change in size with capping enzyme, as expected. (D) Strand specific RT-PCR demonstrates both sense and antisense transcripts for EHI_020920. cDNA was generated using F and R primer (to detect antisense and sense transcript, respectively) as well as oligo dT primer. RT-PCR reveals both antisense and sense transcripts with antisense transcript at lower abundance than sense transcript. Both RT (+) and control reactions lacking RT (−) are shown.

Back to article page