Do I need to get both the First Strand Synthesis Flex Module V2 and the Second Strand Synthesis Module?

No, it depends on what kind of targeted sequencing you intend to do. Both modules are compatible with the QuantSeq FWD V2 (Cat. No. 191 - 196).

Using only the QuantSeq-Flex First Strand Synthesis Module V2 (Cat. No. 166), you can generate target-specific libraries using customized reverse transcription primers with random-primed second strand synthesis. This generates targeted RNA-Seq libraries, allowing the identification of novel fusions.

Using only the QuantSeq-Flex Second Strand Synthesis Module (Cat. No. 028), reverse transcription uses oligo(dT) priming and target-specific primers are used for second strand synthesis. This generates targeted 3' mRNA-Seq libraries.

NOTE: If interested in using QuantSeq-Flex Second Strand Synthesis Module with QuantSeq REV, please contact support@lexogen.com.

Using both the QuantSeq-Flex First and Second Strand Synthesis Modules allows you to generate fully-targeted libraries to identify known targets at specified sequence regions. This is the ideal approach when seeking to replace multiplex qPCR with an amplicon-based targeted RNA-Seq assay.

NOTE: The second strand synthesis reaction requires annealing temperatures of >45 °C. In case you need to use lower temperatures, please contact support@lexogen.com.