No YES. QuantSeq FWD and REV libraries are prepared using the same protocol steps different reagents and reaction volumes.
The use of When Globin or BC1 Blockers Blockers (RS-GBHs / RS-GBSs / RS-BC1B) are added to the preps, the protocol remains unchanged and therefore the same script (QuanSeq QuantSeq FWD or REV) can be applied. This is because blockers are introduced is performed by exchanging these module solutions with the standard RNA Removal Solution (RS) at step 5.
The same applies Ffor users of 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.
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 the first and second strand synthesis steps of library generation. Please contact support@lexogen.com for more information.
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