Overview of sRNA sequencing approach. A Schematic overview of our concatenation procedure. 5' and 3' adapter ligated RNA is reverse transcribed and used as template for PCR. The amplified product is concatenated by blunt ligation in a uncontrolled fashion in which ligation may take place in any direction. The strandedness of the small RNA can still be determined using the original 5' and 3' adapters. B Number of reads corresponding to 1-4 small RNA sequences. 901,887 small RNA reads were obtained from 546,855 sequencing reads, representing a 65% increase in throughput. C Comparison of expression values in the pilot experiment and concatenation experiment. The Pearson correlation coefficient is 0.954, indicating that very little bias is introduced by applying our concatenation strategy. D Frequency distribution of non-redundant sRNA reads with a raw sequence count >1. The number of sequences starting with an A (red), U (green), C (dark blue), and G (cyan) for each size class is indicated.
Klevebring et al. BMC Genomics 2009 10:620 doi:10.1186/1471-2164-10-620