Covaris Adaptive Focused Acoustics® (AFA®) has long been the gold standard for unbiased, efficient, and highly reproducible shearing of DNA for NGS library preparation workflows. The same technology is also available for an unbiased, efficient, and highly reproducible shearing of total RNA, mRNA, and cDNA for RNA-Seq library preparation.
RNA sequencing applications using NGS have been the technique of choice for carrying out whole transcriptome sequencing, monitoring gene expression, and elucidating products of gene splicing and post transcriptional modifications. The recent implication of gene fusion events in tumorigenesis and their potential utility in diagnostics enables a higher level of tumor classification. In drug discovery, it specifically targets products of gene fusion events. This combination brings new focus on the use of RNA-Seq in the clinic for tumor classification, cancer progression, and treatment monitoring.
Chemical fragmentation of RNA is highly sensitive to sample concentration, sample volume, and fragmentation incubation temperature. Physical shearing of RNA using Covaris AFA is concentration independent, isothermal, and highly reproducible. Physical shearing does not require a cleanup step after shearing, thereby preserving the complexity of the sample for library preparation.
Figure 1‐ Agilent BioAnalyzer electropherogram of 100 ng of mRNA processed in a volume of 130 µl of TE in a microTUBE™ according to the operating conditions, showing the mean fragment size at 400 bases.
Figure 2‐ Agilent BioAnalyzer electropherogram of 5 µg of mRNA processed in a volume of 130 µl of TE in a microTUBE according to the operating conditions, showing the mean fragment size at 200 bases.