4. Genotype of sequencing sample.
3. Genotype of Nipponbare. 4. Genotype of R1128. 5. Genotype of sequencing sample.
The authors argued that 95% of the data can be removed in the normal sequencing sample and the percentage reaches 98% of high-coverage single sequencing data.
Overall, we identified 575 circRNA species that pass a beta-binomial test for enrichment (false discovery rate of 1%) in the exonuclease-treated sequencing sample.
Approximately 30 million reads were generated from each sequencing sample, of which 58.10% 65.42% could be mapped to the C. elegans genome (WS190) with two or less mismatches (Table S4).
Procedures to improve on library preparation for sequencing sample preparation have also been published [7], [8], [9], [10].
These results suggest that considerable improvements in absolute base coverage may be achieved through relatively simple and cost-effective modifications of high-throughput sequencing sample preparation protocols.
Each sequencing sample contained 20 µL of de-ionized, DNA-free water (Invitogen), and 4 µL of the diluted ssDNA product in Tris HCl Buffer pH 8.0 (Sigma).
One site each in beaver samples C. canadensis and C. fiber albicus had less than 3x coverage, for which the sequence was confirmed by PCR and Sanger sequencing (sample specific primers listed in Table S8).
Each air-dried sequencing sample was dissolved in 10 µl Hi-Deionized Formamide (Applied Biosystems Inc., Foster City, California), denatured at 95°C for 1 min and immediately held at 4°C prior to sequencing with a Model 3100 Genetic Analyzer (Applied Biosystems Inc., Foster City, California).
cDNA libraries were prepared according to Illumina sequencing sample preparation protocols.
