Deep Profiling of the Novel Intermediate-Size Noncoding RNAs in Intraerythrocytic Plasmodium falciparum
Figure 4
Experimental validation of novel is-ncRNAs.
(A) RT-PCR confirmation of the novel is-ncRNAs. Twelve of the is-ncRNAs are shown as examples, and the remaining 48 are documented in Figure S1 in file S2. Each is-ncRNA is shown in three adjacent lanes, from left to right: DNase-treated RNA RT-PCR (“+”), RT-PCR with no RNA template (left, “−”, negative control), and RT without reverse transcriptase (right, “−”, negative control). “M” indicates the 50 bp DNA ladder. “Length” indicates the expected size of the is-ncRNAs based on the Illumina/Solexa paired-end sequencing assembly data. (B) Northern blotting validation of six novel is-ncRNAs. U5 snRNA was used as an internal control. The arrows indicate the sizes of the is-ncRNAs; the black arrows indicate products with lengths consistent with those of the RT-PCR products, and the grey arrows indicate products with lengths that were larger than those determined by RT-PCR. “Length” indicates the expected size of the is-ncRNA based on the Illumina/Solexa paired-end sequencing assembly data and RT-PCR.