FAQ: Is the Monarch® Total RNA Miniprep Kit compatible with NEBNext® reagents for RNA Library Prep?

Yes. RNA purified with this kit can be used for RNA library preparation using NEB's NEBNext RNA Library Prep reagents.


Monarch-purified RNA can be used to prepare high quality RNA-seq libraries for gene expression analysis.

RNA Sequence
Transcript levels in Universal Human Reference RNA (UHRR, Agilent) are compared before and after re-purification using either Qiagen RNeasy® or the Monarch Total RNA Miniprep Kit. Strong correlation with untreated UHRR is observed for both methods (Pearson R > 0.99 for both samples). All samples display consistent end-to-end coverage of transcripts indicating an absence of detectable degradation during purification. Poly-A selected RNA was selected from 100 ng of untreated, Qiagen and Monarch samples using the NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490). RNA-seq libraries were then prepared using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina® (NEB #E7760) before sequencing on a Miseq® instrument (2 x150). 1.6M reads were randomly sampled from each library and adaptor trimmed (Seqprep v1.1). Levels of all Gencode v26 transcripts were assessed using Salmon (0.4) and plotted above (panel A). Average 5'-3' Coverage of Gencode v26 transcripts (assessed by Picard's CollectRnaSeqMetrics 1.56 after mapping to the GRCh38 reference genome with Hisat v2.0.3 and marking duplicates with Picard's MarkDuplicates 1.56 ) is shown below (panel B).