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

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) is performed by exchanging these module solutions with the standard RNA Removal Solution (RS) at step 5.

For users of the UMI Module (USS), the Second Strand Synthesis Mix 1 (SS1) 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. Please contact support@lexogen.com for more information.