True and accurate small RNA-seq:
Use the power of UMIs without ligation
D-Plex Small RNA-seq library preparation solution
Diagenode’s latest innovation in small RNA-seq, D-Plex,is based on a template-switching technology that delivers
a ligation-free method for RNA library preparation,
ensuring realistic representation of diverse small RNA
species (such as miRNA, piRNA, tRNA, and siRNA) withminimized bias. This solution allows you to produce
Illumina®-compatible DNA libraries for NGS from low
input amounts of RNA.
The D-Plex Small RNA Small solution was optimized for10 pg - 10 ng enriched small RNA (or 100 pg - 100 ng
total RNA) and contains our latest TSO (Template-SwitchOligonucleotide) technology including UMIs.

Benefits of D-Plex: Truest representation: UMIs with ligation-freeprotocol ensures the most representative andunbiased data
 Enhanced data: Our new bioinformatics pipeline
provided in the manual was optimized and developed
specifically for getting the best out of your D-Plexdata including UMI processing Easy: One-tube, one-day protocol
 owest inputs: Down to 10 pg enriched small RNALor 100 pg total RNA starting amount Clinical sample compatibility: Tested with RNA frombiofluids

D-Plex provides high transcript diversity

Figure 1. Biotyping : Relative
abundance of each small RNA
from Arabidopsis, Human andMouse type captured in the libraryout of the total amount of RNA.
(RNA transcripts annotated inEnsembl).

1 ng 100 ng 100 pg 1 ng 100 ng 100 pg
Leaves

Roots

1 ng

10 pg 100 pg 1 ng

10 pg 100 pg

PlasmaSerum

10 ng 10 pg 100 pg 1 ng 100 ng 100 pg
Small RNA

Total RNAHow D-Plex works

RNASTEP
2AAAAA...

RNA
UMI+5’ adaptor

STEP
3rXrXrXrX

RNA sample
preparation

20 minRNA dephosphorylationand tailing

1h30

Reverse transcription>2h

TPM - 10 pg

RNASTEP
1Robust technology no matter your input amountTime Pause

AAAAAAA...V TTTTTTTT...XXXXAAAAAAA
TTTTTTTT

XXXX
XXXX

AAAAAAA
TTTTTTTTPCR forward
PCR reverse

STEP
4

TPM - 10 ng

AAAAAAA
TTTTTTTT

XXXX
XXXX

PCR amplification

45 minFigure 3. Correlation between 2 different starting amounts
using D-Plex Small RNA-seq Kit. 10 ng and 10 pg of small RNA
were isolated for ncRNA detection at TPM ≥2. Blue line: idealcorrelation; Red line: r = 0.97.

DNAFigure 2. D-Plex one-day workflow.
STEP
5

Library purificationand shaping

UMIs allow accurate identification ofduplicates for low input samples
1 ng

High read quality with D-Plex Small RNA

10 pg

1_ng

10_pg

STEP
6

Ensembl transcript biotype
miRNA
misc_RNAribozyme
scaRNA
snoRNA

10.0

snRNA

1.00.1

1

10100

1000

10000

1Before UMI deduplication

101001000

10000

Position in read (bp)log10 countFigure 4. Effect of the UMI deduplication on the sncRNA set
detected at TPM � 2 in rat plasma for different inputs: 1 ng
and 10 pg. (RNA transcripts annotated in Ensembl).

Figure 5. D-Plex validated on NovaSeq 6000 (100 bp SE reads)
using 100 pg RNA isolated from human serum (PhiX = 3%).

Description

Cat. No.

Format

D-Plex Small RNA-seq Kit*

C05030001

24 rxns

D-Plex 24 Single Indexes for Illumina - Set #A (1-24)

C05030010

24 rxns

D-Plex 24 Single Indexes for Illumina - Set #B (25-48)

C05030011

24 rxns*does not contain indexes

www.diagenode.com Please contact us for more information
Europe Diagenode sa / LIEGE SCIENCE PARK // Rue Bois Saint-Jean, 3 // 4102 Seraing (Ougrée) // Belgium // Phone: (+32) 4 364 20 50 // E-mail: info@diagenode.com
USA Diagenode Inc. / 400 Morris Avenue, Suite 101 // Denville, NJ 07834 // USA // Phone: (+1) 862 209-4680 // E-mail: info.na@diagenode.com

F-D-plex-01-20

log10 countAfter UMI deduplication

100.0

Phred score

Library quality control
and NGS

1000.0