Is there a separate automation protocol for QuantSeq REV library preps or for libraries prepared with Globin or BC1 Block and/or UMIs?

YES. QuantSeq FWD and REV libraries are using different reagents and reaction volumes.

When Globin or BC1 Blockers (RS-GBHs / RS-GBSs / RS-BC1B) are added to the preps, the protocol remains unchanged and therefore the same script (QuantSeq FWD or REV) can be applied. This is because blockers are introduced by exchanging these module solutions with the standard RNA Removal Solution (RS) at step 5.

The same applies for the UMI Module (USS), where the Second Strand Synthesis Mix 1 (SS1) is exchanged at step 7 of the QuantSeq FWD protocol. The UMI module is not compatible with the QuantSeq REV kit.

NOTE: QuantSeq Flex libraries may require different automated protocol modifications depending on the use of custom primers at either or both of the first and second strand synthesis steps of library generation. Please contact support@lexogen.com for more information.