Examples of nanoblots, comparison to northern blots, overlaying, size estimation, and conditional probes. (A) Comparison between 32P northern Blot and nanoBlot targeting the 5′-exon of RPL18A. Notice the accumulation of the small molecular weight species in the rrp6Δ samples. S: Spliced RPL18A mRNA, CF: 5′ cleavage fragment. (B) Example of nanoblot on the low abundance gene, ADI1. Also shown is a nanoblot with the size axis shown on a log2 scale. (C) Overlaid nanoblot showing sequencing reads obtained from two genes probed with different colors: RPS7B = orange; snR4 = cyan. (D) The same data as in C but with each probe + samples combination separated into its own lane. (E) Nanoblot showing size misrepresentation of snR37. The red line indicates the true size of the mature snoRNA. slu7-AA: Direct RNA sequencing of Slu7-FRB rapamycin-treated RNAs preprocessed with Terminator exonuclease, rSAP, and in vitro polyadenylated. (F) IGV screenshots showing the reads selected by the use of a single probe (shown in green) (left) or a combination of one probe and an antiprobe (shown in red) (right). (G) Nanoblot produced for the data shown in F using the combination of probe only (−) or probe and antiprobe (+).
