The REPLI-g WTA Single Cell Kit enables reliable investigation of effects on transcription regulation at the single-cell transcriptome level and allows uniform amplification of all transcripts from just single cells (1–1000 cells). Dedicated buffers and reagents undergo a unique, controlled decontamination procedure to block amplification of contaminating nucleic acids by the REPLI-g method. The innovative lysis buffer effectively stabilizes cellular RNA, ensuring the resulting RNA accurately reflects the in vivo gene expression profile. All enzymatic steps have been developed to enable efficient processing of RNA for accurate amplification of cDNA, which is achieved with negligible sequence bias using innovative Multiple Displacement Amplification (MDA) technology

1. Sample to Insight
Whole transcriptome amplification (WTA) from single cell
Learn WTA of total RNA or mRNA from single cells

2. Sample to Insight
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Discover the REPLI-g WTA Single Cell Kit
• High dynamic range of transcripts
◦ MDA-based WTA technology without length bias
• Superior sensitivity
◦ Low-abundance transcript detection
• Superior sequence accuracy
◦ More confidence in your data in sensitive applications like viral RNA analysis
• For downstream NGS, arrays, aCGH and PCR
◦ Free choice of downstream analysis
• Enables bio-banking
◦ Amplified cDNA can be stored for follow-up studies or confirmatory testing.
PCR-free WTA
technology
Poly-A or total
RNA protocol
Start from a single
eukaryotic cell
*MDA = multiple displacement amplification
Single cell genomics by QIAGEN, 2016

3. Sample to Insight
For single cell WTA
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Ideally suited for
• Sensitive transcript discovery and differential gene
expression analyses from single eukaryotic cells
• Downstream analysis using aCGH, PCR or NGS
• Multiple analyses from a single cell
• Researching infectious diseases, viral RNA enricment and
analysis
• Single cell RT-PCR analyses, event confirmation in genome
engineering applications
• Bio-banking the RNA content of a single cell
REPLI-g WTA
Single Cell Kit
Single cell genomics by QIAGEN, 2016

4. Sample to Insight
REPLI-g WTA Single Cell Kit
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WTA for sensitive applications (e.g. single cell qRT-PCR/viral RNA analyses
Primary
sample
isolation
Single cell
isolation WTASample
• Single eukaryotic cells
• Picogram levels of
purified DNA
• Free choice of downstream analysis
◦ NGS
◦ Microarray
◦ PCR
• Multiple analyses from a single cell
InsightPCR
Data
analysis
Interpretation
NGS
Microarray
Single cell genomics by QIAGEN, 2016

5. Sample to Insight
REPLI-g WTA Single Cell Kit
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More reliable detection of even low-
abundance transcripts.
Whole transcriptome amplification was
performed using 20 pg of total RNA.
Unlike kits from other suppliers, which
were less successful at amplifying the same
amount of the same transcript, the REPLI-g
WTA Single Cell Kit enabled detection of
100% of high-, medium- and low-abundance
transcripts following RNA amplification.
Real-time PCR of 22 medium-abundance
transcripts, representing the medium-
abundance transcript row in [A],
demonstrated that only the REPLI-g
WTA Single Cell Kit reliably amplified all
transcripts. Transcripts that could not be
detected gave CT values >40 (red bar).
Reliable detection of even low abundance transcripts
Single cell genomics by QIAGEN, 2016

6. Sample to Insight
REPLI-g WTA Single Cell Kit: key citations
6
(1) Lee, D. et al. (2015) Interplay between glucose and leptin signaling determines the strength of GABAergic synapses at POMC neurons. Nat. Commun. 26:6, 6618
(2) Jeong, J.H. (2015) Cholinergic neurons in the dorsomedial hypothalamus regulate mouse brown adipose tissue metabolism, MOLECULAR METABOLISM 4
(3) Lee, D. et al. (2015) Apelin-13 Enhances Arcuate POMC Neuron Activity via Inhibiting M-Curren. PLoS ONE 10(3): e0119457.doi:10.1371/journal.pone.0119457
(4) Chen, J. et al. (2015) Heterodimerization of Human Orexin Receptor 1 and Kappa Opioid Receptor Promotes Protein Kinase A/cAMP-Response Element Binding
Protein Signaling via a Gαs-Mediated Mechanism. Cell. Signal., 27, 1226–1438.
Gene expression
analysis by single
cell qRT-PCR
• Single cell qRT-PCR was used to measure the abundance of transcripts involved in leptin
signaling to help build a model that integrates both glucose and leptin signaling pathways in
proopiomelanocortin (POMC) neurons. (1)
• RT-PCR analysis revealed the co-expression of muscarinic
acetylcholine receptor (M2 mAChR) and tryptophan hydroxylase 2
(Tph2) in single cells collected from mouse brain slices. (2)
• In this study, single cell qRT-PCR was used to monitor changes in the
expression of phospholipase C splice variants involved in signaling. (3)
• Single cell analysis was applied to identify co-expression of orexin receptor 1 (OX1R) and
kappa opioid receptors (KOR) in rat hippocampal neurons. (4)
Single cell whole transcriptome amplification (WTA) with REPLI-g WTA Single
Cell Kit, followed by qPCR analysis
Single cell genomics by QIAGEN, 2016

7. Sample to Insight
REPLI-g WTA Single Cell Kit: key citations
7
Proof-of-concept study: detection & extended characterization of
Norovirus genotypes by NGS from norovirus positive fecal
samples
(1) Bavelaar, H.H. (2015) Whole genome sequencing of fecal samples as a tool for the diagnosis and genetic characterization of norovirus. J. Clin. Virol. 72, 122
• Whole transcriptome amplification of RNA with REPLI-g WTA Single
Cell Kit
• NGS, followed by data analysis on CLC Genomics Workbench
Virus
research
Single cell genomics by QIAGEN, 2016

8. Sample to Insight
Single cell genomics by QIAGEN
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For in-depth, molecular analysis of single cells
Single cell
WGA or WTA
Single cell
isolation
Single cell
sequencing
• Easily access
single cells
• Affordable
• Gentle on cells
QIAscout
QIAseq
FX Single Cell
Library Kits
REPLI-g
Single Cell Kits
Single cell
miRNA analysis
• Create superior-
quality NGS
libraries directly
from single cells
• Provides best-in-
class amplification
of genomes or
transcriptomes from
single cells
• Profile miRNA
expression using
qPCR
miScript
Single Cell
qPCR Kit
Single cell genomics by QIAGEN, 2016
Visit the single cell resource center