No. QuantSeq FWD and REV libraries are prepared using the same protocol steps and reaction volumes.
The use of Globin or BC1 Block (RS-GBHs / RS-GBSs / RS-BC1B) or the UMI Module (USS) , is performed by exchanging these module solutions with the standard RNA Removal Solution (RS) , or at step 5.
For users of the UMI Module (USS), the Second Strand Synthesis Mix 1 (SS1) at the respective steps (5 and 7). The is exchanged at step 7.
For both modules, the transferred volume is the same at each step so only the supplied reagent tube needs to be exchanged – no protocol or volume changes are required.
Therefore, the same automation protocol can be used for preparing all types of QuantSeq FWD and REV libraries, with or without Globin Block and / or UMIs.
NOTE: QuantSeq Flex libraries may require different automated protocol modifications depending on the use of custom primers at either or both of first and second strand synthesis steps of library generation. QuantSeq Flex programming is available for the Biomek i7 and NGS Workstation B (Agilent Bravo). Please contact support@lexogen.com if you plan on automating QuantSeq Flex on any liquid handling platforms for more information.