This document discusses single cell genomics and describes QIAGEN kits that can be used to analyze individual genomes from single cells. The kits use REPLI-g technology to overcome typical challenges in whole genome amplification and whole transcriptome amplification from single cells, such as low and uneven coverage and allelic dropout. The technology provides high yields and uniform coverage with low base copying errors compared to other methods like MALBAC. The kits allow characterization of individual genomes without cell culturing and have applications in cancer research, microbial analysis, and studying cellular heterogeneity.

1. Get the most from a single cell
Analyze cellular
heterogeneity during
immune or stem cell
development
Explore tumor
heterogeneity to
understand cancer
evolution and subclonal
architecture
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A T C G G A T C A T C G G A T
A T C G G A T C A T C G G A T C A
A T C G G A T C A T C G G A T C A T C
A T C G G A T C A T C G G A T C A T C G G
A T C G G A T C A T C G G A T C A T C G G A T
A T C G G A T C A T C G G A T C A T C G G A T C A
A T C G G A T C A T C G G A T C A T C G G A T C A T C
A T C G G A T C A T C G G A T C A T C G G A T C A T C G G
A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T
A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T
A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T C G
A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T C G G A
A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T C G G A T C
A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T C G G A T C A T
A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T C G G A T C A T C G
A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T C G G A T C A T C G G A
A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T C G G A T C A T C G G A T A
A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T C G G A T C A T C G G A T A T C
A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T C G G A T C A T C G G A T A T C G G
T C A T C G G A T C A T C G G A T A T C G G A T C A T C G G A T A T C G G A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T C G G A T C A T C G G A T A T C G G A T C G G A T C A T C G G A T C A T
A T C A T C G G A T C A T C G G A T A T C G G A T C A T C G G A T A T C G G A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T C G G A T C A T C G G A T A T C G G A T C G G A T C A T C G G A T C AT C G G A T C A T C G G A T C A T C G G A T A T C G G A T C A T C G G A T A T C G G A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T C G G A T C A T C G G A T A T C G G A T C G G A T C A T C G GA T C G G A T C A T C G G A T A T C G G A T C A T C G G A T A T C G G A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T C G G A T C A T C G G A T A T C G G A T C G G A T C A T C G G A T C A T C GG A T C A T C G G A T C A T C G G A T A T C G G A T C A T C G G A T A T C G G A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T C G G A T C A T C G G A T A T C G G A T C G G A T C A T C G G A T CC G G A T C A T C G G A T A T C G G A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T C G G A T C A T C G G A T A T C G G A T C G G A T C A T C G GC G G A T C A T C G G A T A T C G G A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T C G G A T C A T C G G A T A T C G G A T C G G A T C A T C G
A T C G G A T C A T C
A T C G G A T C A T C G G
A T C G G A T C A T C G G A T
A T C G G A T C A T C G G A T C A
A T C G G A T C A T C G G A T C A T C
A T C G G A T C A T C G G A T C A T C G G
A T C G G A T C A T C G G A T C A T C G G A T
A T C G G A T C A T C G G A T C A T C G G A T C A
A T C G G A T C A T C G G A T C A T C G G A T C A T C
A T C G G A T C A T C G G A T C A T C G G A T C A T C G G
A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T
A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T
A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T C G
A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T C G G A
A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T C G G A T C
A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T C G G A T C A T
A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T C G G A T C A T C G
A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T C G G A T C A T C G G A
A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T C G G A T C A T C G G A T A
A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T C G G A T C A T C G G A T A T C
A T C G G A T C A T C G G A T C A T C G G A T C A T C G G A T A T C G G A T C A T C G G A T A T C G G
Characterize individual
genomes and eliminate
the need for culturing
Characterize the
genome and
transcriptome of CTCs –
applications for liquid
biopsy
Circulating
tumor cells (CTCs)
Single cell
genomics
Microbial
samples
Cancer
research
Typical challenges in
WGA + WTA
REPLI-g®
MDA technology for overcoming challenges
Get the most from a single cell with these QIAGEN kits:
1. High enzyme processivity – no dissociation, pausing or slippage – long reads (>70 kb)
2. Superior proofreading activity with high-fidelity enzyme – 1000-fold higher fidelity than normal PCR enzymes (1)
3. High yields – get sufficient material for your downstream applications, including NGS, PCR or microarrays
4. Optimized lysis and DNA denaturation – immediate amplification across all regions
(1) J.Liang et al., Journal of Genetics and Genomics 41 (2014) 513-528
Sample to Insight
Coverage
uniformity
Allelic
drop-out
T • • G T •
Amplification
factor/yield
Base
copying error
T • G •
— REPLI-g Single Cell Kit
— REPLI-g WTA Single Cell Kit
— REPLI-g Cell WGA & WTA Kit
— REPLI-g Single Cell DNA Library Kit
— REPLI-g Single Cell RNA Library Kit
+ NGS library prep
+ NGS library prep
Trademarks: QIAGEN®
, Sample to Insight®
, REPLI-g®
(QIAGEN Group). For up-to-date licensing information and product-specific disclaimers, see the respective kit handbook or user manual.
© 2015 QIAGEN, all rights reserved. PROM-8288-001.
50 µg
0 µg
REPLI-g MALBAC
46
3
Amplification yield
1%
0 %
REPLI-g MALBAC
0.2
Error rate
0.7
REPLI-g generates >10x the yield with
significantly better accuracy than MALBAC
REPLI-g provides more
uniform coverage
REPLI-g
MALBAC
Normalized coverage
1.4
1.2
1.0
0.8
0.6
0.4
0.2
0
20
GC content (%)
30 40 50 60 70
1 pg DH10B DNA, amplified with either REPLI-g Single Cell Kit or by MALBAC, sequenced on MiSeq Illumina (V2, 2x150 nt).