What are the differences between QS FFPE and QuantSeq V2 kits?

QuantSeq FFPE

QuantSeq V2*

First strand cDNA synthesis

• RNA input volume up to 7.5µl

• Oligod(T) with UMIs

• Oligo dT provided separately

• Denaturation step 1min at 95°C

• 10X FS buffer

• RT reaction 15min

• RNA input volume up to 5µl

• No UMIs included

• Oligod(T) primer included in the FS1 buffer

• Skip denaturation step

• FS1 and FS2 buffers

• RT reaction up to 1hr

RNA removal

• RS + RE enzyme

• 3 µl Reaction volume of RS + RE mastermix added per reaction

• Additional incubation step at 37°C, prior to incubation at 95°C

• Only RS enzyme

• 5µl reaction volume of RS added per reaction

• Incubation at 95°C only

Second Strand Synthesis

• 5µl SS1 buffer

• 15 min incubation

• 2µl SS2/E2 reaction mix

• 10µl SS1 buffer

• 30 min incubation

• 5µl SS2/E2 reaction mix

Purification

• 8µl PB

• 20µl EB

• 26µl PS

• 16µl PB

• 40µl EB

• 56µl PS