Capture and Amplification by Tailing and Switching (CATS) method has been previously developed by scientists working at the German Cancer Research Center and SciBerg e.Kfm, and allows efficient deep sequencing of RNA, DNA and bisulfite-converted DNA from ultra-low inputs, including nucleic acids obtained from biological fluids (liquid biopsies). CATS technology can have broad applications in various fields, including translational medicine for therapy monitoring, prediction, prognosis and early detection of various human disorders. For more information please watch the recent seminar held by a SciBerg founder, Dr. Turchinovich, as a part of the LabRoots 5th Annual Genetics and Genomics virtual conference.
Fragmented (3 - 600 nt long) single-stranded RNA, DNA or bisulfite-converted DNA is polyadenylated or polydeoxyadenylated and converted into cDNA by MMLV reverse transcriptase in the presence of the anchored poly(dT) primer containing a custom terminal adaptor sequence. After the MMLV reverse transcriptase reaches the 5′ end of the template, it switches the template into the so-called, template switching oligonucleotide (TSO) which serves as a second template for the reverse transcriptase. TSO consists of three 3′-terminal rG nucleotides and a custom terminal adaptor sequence. The second cDNA strand is generated during the first cycle of the final pre-amplifying PCR reaction. Other things being equal, NGS libraries produced by CATS approach have significantly higher complexity as compared to the libraries generated by ligation-based methods. Indeed, polynucleotide tailing is a much more efficient way to capture 3-OH termini of single-stranded nucleic acids than ligation of adapters and also have little or no biases. Besides, CATS require much lower inputs of nucleic acids and can prepare NGS libraries from picogram inputs of highly fragmented RNA or DNA. Finally, CATS is capable to efficiently capture highly fragmented single-stranded DNA molecules (e.g. bisulfite-converted DNA, highly degraded DNA, etc.).
CATS RNA-seq & DNA-seq kits
Diagenode SA has recently acquired the exclusive worldwide license for the CATS technology and is offering a complete portfolio of NGS library preparation kits optimized for specific research applications. For more information please visit Diagenode website. For information about licensing of CATS method with respect to diagnostics and therapeutics, please contact Technology Transfer Heidelberg GmbH.