Lexogen’s QuantSeq-Flex kit provides a protocol to generate Illumina-compatible libraries from total RNA within 4.5 hours. The available flexible modules for First Strand and/or Second Strand cDNA Synthesis allow the use of custom primers and renders the kit suitable for targeted sequencing. The QuantSeq-Flex protocol generates only one fragment per transcript, resulting in extremely accurate gene expression values, regardless if oligodT priming or target specific priming is used during first strand synthesis.
The kit upgrade to QuantSeq-Flex V2 brings a new, streamlined protocol for target-specific second strand synthesis. RNA hydrolysis is now integrated into the second strand synthesis, saving time and effort, and the annealing of targeted primers can now be at 45 – 72 °C, increasing specificity and making primer design more flexible. Safe stopping points are indicated, and the QuantSeq-Flex First Strand Synthesis Module is now recommended to generate QuantSeq 3‘ mRNA-Seq libraries with increased insert sizes for sequencing runs with longer reads.
These upgrades further increase the applicability of QuantSeq-Flex for targeted RNA-Seq, for amplicon sequencing as well as for targeted 3’ mRNA-Seq. The kit is also ideal for customization, as shown by the Ameres group, which generated 3’ RNA-Seq libraries from immunoprecipitated (nc)RNAs 3’ ligated to custom linkers: Reimao-Pinto, et al. (2016) Molecular basis for cytoplasmic RNA surveillance by uridylation‐triggered decay in Drosophila. The EMBO Journal: e201695164
Learn more about QuantSeq-Flex on its web page.