What are the updates to QuantSeq REV V2?

The QuantSeq 3‘ mRNA-Seq V2 Library Prep Kit REV with Unique Dual Indices (12nt, Cat. No. 225) replaces the previous version of the QuantSeq 3‘ mRNA-Seq Library Prep Kit REV (Cat. No. 016).

In the V2 kits, there are several component changes:

• First Stand cDNA Synthesis Mix “FS1” and First Strand cDNA Synthesis Mix 2 “FS2” are replaced with 10x First Strand cDNA Synthesis Mix “10xFS”, OligodT primer REV “dT REV”, and “DTT”

• PCR Mix “PCR” and Enzyme Mix 3 “E3” are replaced with PCR Mix “PM” and PCR Enzyme Mix “PE”

• Lexogen i7 6nt Index Set is replaced with Lexogen UDI 12nt Set B

Along with the update to our PCR components, there was an update to the PCR program as shown below:

• PCR program of the QuantSeq REV with UDI V1 library amplification:

• PCR program of the QuantSeq REV with UDI V2 library amplification:

The QuantSeq REV V2 Kits allow for lower RNA input amounts, down to 1 ng of total RNA. A modified protocol, included in the QuantSeq REV V2 User Guide (Cat.No.225), can be used for such low inputs. Finally, higher RNA input volumes, up to 16 μl, can be used.