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Unless otherwise stated in references, presented data on file For Research Use Only. Not for use in diagnostic procedures.
Shift your workflow into hyperdrive.KAPA HYPER PREP KITS
The KAPA Hyper Prep Kit is a versatile, streamlined library prep solution that provides higher yields of adapter-ligated libraries and lower amplification bias enabling:
• superior speed and convenience
• lower duplication rates and higher sequencing coverage
• improved performance with low-input samples
• high-quality library construction from FFPE samples
• PCR-free workflows
KAPA Hyper Prep Kits | 2For Research Use Only. Not for use in diagnostic procedures.
Unless otherwise stated in references, presented data on file
% C
on
vers
ion
DNA input
0
5
10
15
20
25
30
35
40
1 µg 100 ng 10 ng
Superior Speed and Convenience
High Library Yields and Sequencing Quality
The streamlined, one-tube KAPA Hyper Prep
protocol offers the fastest turnaround time,
with minimal hands-on time.
• Complete library construction in less
than 3 hours
• Library amplification may be omitted
for PCR-free workflows
• Bead-based size selection steps can be
incorporated to achieve the appropriate
final library fragment-size distribution
• Achieve higher library yields across a
range of input DNA and sample type
• Reduce the number of amplification cycles
needed for downstream processing, resulting
in lower duplication rates and higher
sequence coverage
• Enable successful library construction for
challenging samples and PCR-free workflows
KAPA Hyper Prep Kit
Total time: 2.75 hours
End Repair & A-tailing
Adapter Ligation
Library Amplification
Bead Cleanup
Bead Cleanup
NEBNext® Ultra Library Prep Kit
Total time: 3 hours
End Repair & A-tailing
Adapter Ligation
USER Excision
Library Amplification
Bead Cleanup
Bead Cleanup
Illumina® TruSeq® Nano DNA Sample Prep Kit
Total time: 6 hours
End Repair
A-tailing
Adapter Ligation
Library Amplification
Bead Cleanup
Size Selection
Bead Cleanup
Conversion rates for libraries prepared for target capture from different amounts of Covaris-sheared DNA, using the KAPA Hyper Prep or competitor library construction kits. Libraries were prepared according to manufacturer’s instructions. Input DNA was quantified by Qubit, whereas the qPCR-based KAPA Library Quantification Kit was used to determine KAPA Hyper Prep and TruSeq Nano library yields after adapter ligation. Conversion rates for NEBNext libraries cannot be measured directly using the KAPA Library Quantification Kit and were derived from post-amplification yields.
Conversion Rate Comparison
KAPA Hyper Prep
TruSeq Nano DNA Kits
NEBNext Ultra
% C
onve
rsio
n
DNA Input
Conversion rate, defined as % input DNA converted to sequenceable,
adapter-ligated library, is a key library construction metric which
ultimately determines library diversity and quality.
KAPA Hyper Prep Kits | 3For Research Use Only. Not for use in diagnostic procedures.
Unless otherwise stated in references, presented data on file
125.3X
97.5 96.2
87.7
56.4
76.1X
97.1 94.2
71.3
24.6
0
25
50
75
100
125
150
Mean Depth % Bases at 10X
% Bases at 20X
% Bases at 30X
% Bases at 50X
% Bases at 100X
94.4
89.2
Sequence Coverage by Depth
KAPA Hyper Prep
TruSeq® Nano DNA Kits
High-quality Library Construction from FFPE Samples
• Detect low-frequency mutations with lower
duplication rates and high sequence coverage
• Improve library diversity with more unique
adapter-ligated library fragments from
low- quality sample types
19.5
6.4
0
5
10
15
20
25
KAPA Hyper Prep
TruSeq Nano DNA
Library Diversity (108 molecules)
KAPA Hyper Prep
TruSeq Nano DNA
9.7
19.5
0
5
10
15
20
25
Duplication (%)Sequencing metrics for libraries prepared from 100 ng FFPE DNA for target capture, using either the KAPA Hyper Prep Kit or TruSeq® Nano DNA Sample Prep Kit (Illumina). Captures were performed with the SeqCap™ EZ Comprehensive Cancer Design (4 Mb; Roche® NimbleGen™) according to the manufacturer’s instructions, with the exception that the number of pre-capture amplification cycles for each library type was optimized based on post-ligation yields (10 cycles for KAPA Hyper Prep vs. 14 cycles for TruSeq Nano DNA libraries). All libraries were amplified for 13 cycles after capture. Sequencing (2 x 75 bp) was performed on an Illumina HiSeq®. Sequencing reads were down-sampled to ~14 million per library prior to analysis with Picard.
Library Diversity Duplication Rates
x108 m
olec
ules
(%)
Minimal Amplification Bias
• In workflows where amplification is
required, KAPA HiFi introduces minimal
amplification bias, resulting in more
uniform sequence coverage
• Kits available with amplification module
or for PCR-free workflows
GC bias plots for libraries prepared for whole-genome shotgun sequencing of bacteria with extreme genomic GC content. Libraries were prepared with the KAPA Hyper Prep Kit from 100 ng or 1 ng DNA, Covaris-sheared to an average size of ~200 bp, and sequenced without amplification, or after amplification with KAPA HiFi. Sequencing (2 x 300 bp) was performed on an Illumina® MiSeq®, and data analyzed using Picard.
Normalized Coverage
Windows at GC%
Base Quality at GC%
Bordetella_100ng_Covarls_NoClean_Hyper_NoAmp.bam.null GC Blas PlotTotal clusters: 319,787, Aligned reads: 600,235
0 20 40 60 80 100
0.0
0.5
1.0
1.5
2.0
GC% of 100 base windows
Fra
ctio
n o
f no
rmal
ized
cov
erag
e
01
02
03
04
0
Mea
n b
ase
qu
alit
y
0 20 40 60 80 100
0.0
0.5
1.0
1.5
2.0
GC% of 100 base windows
Fra
ctio
n o
f no
rmal
ized
cov
erag
e
01
02
03
04
0
Bordetella_1ng_Covaris_NoClean_Hyper_Amp.bam.null GC Bias PlotTotal clusters: 312,391, Aligned reads 580,794
Mea
n B
ase
Qu
alit
y
Total Clusters: 319,787Aligned Reads: 600,235
Total Clusters: 312,391Aligned Reads: 508,794
100 ng PCR-free 1 ng amplified
Bor
dete
llaFr
acti
on o
f nor
mal
ized
cov
erag
e
GC% of 100 Base Windows GC% of 100 Base Windows
Frac
tion
of n
orm
aliz
ed c
over
age
Mea
n B
ase
Qua
lity
Mea
n B
ase
Qua
lity
Clostridium_100ng_Covaris_NoClean_Hyper_NoAmp.bam.null GC Bias Plot Total clusters: 239,377, Aligned reads: 458,669
GC% of 100 base windows
Fra
ctio
n o
f no
rmal
ized
cov
erag
e
Mea
n b
ase
qu
alit
y
0 20 40 60 80 100
0.0
0.5
1.0
1.5
2.0
01
02
03
04
0
Clostridium_1ng_Covaris_NoClean_Hyper_Amp.bam.null GC Bias Plot Total clusters: 233,138, Aligned reads: 444,053
GC% of 100 base windows
Fra
ctio
n o
f no
rmal
ized
cov
erag
e
Mea
n b
ase
qu
alit
y
0 20 40 60 80 100
0.0
0.5
1.0
1.5
2.0
01
02
03
04
0
Total Clusters: 239,377Aligned Reads: 458,669
Total Clusters: 233,138Aligned Reads: 444,053
Clo
stri
dium
GC% of 100 Base Windows GC% of 100 Base Windows
Frac
tion
of n
orm
aliz
ed c
over
age
Mea
n B
ase
Qua
lity
Frac
tion
of n
orm
aliz
ed c
over
age
Mea
n B
ase
Qua
lity
Ordering Information
Improved Performance with Low-input Samples
• Generate more diverse libraries from limited amounts of input DNA
• High adapter:insert molar ratios can be used to increase library
construction efficiency
Library diversity and duplication rates for libraries prepared from 2 ng cell-free DNA, using the KAPA Hyper Prep Kit or a leading competitor kit optimized for low-input library construction. Libraries were prepared according to manufacturer’s instructions using standard ligation parameters. KAPA Hyper Prep libraries were prepared with a range of adapter:insert molar ratios. Sequencing (2 x 150 bp) was performed on an Illumina® MiSeq®, and data analyzed using Picard.
Duplication RatesLibrary Diversity
x108 m
olec
ules
(%)
0
1
2
3
1.52
Hyper (60:1)
1.74
Hyper (300:1)
2.64
Hyper (1000:1)
1.85
Competitor
Library Diversity (108 molecules) Duplication Rates
0.05
0.10
0.15
0.20
0.25
0.00
0.21
Hyper (60:1)
0.18
Hyper (300:1)
0.17
Hyper (1000:1)
0.18
Competitor
KapaBiosystemsKapaBiosystems@KapaBiosystems
Roche Cat. No. Kapa Code Description Kit Size
07962312001 KK8500 KAPA Hyper Prep Kit with Library Amplification 8 reactions
07962347001 KK8502 KAPA Hyper Prep Kit with Library Amplification 24 reactions
07962363001 KK8504 KAPA Hyper Prep Kit with Library Amplification 96 reactions
07962339001 KK8501 KAPA Hyper Prep Kit, PCR-free 8 reactions
07962355001 KK8503 KAPA Hyper Prep Kit, PCR-free 24 reactions
07962371001 KK8505 KAPA Hyper Prep Kit, PCR-free 96 reactions
Unless otherwise stated in references, presented data on file© 2016 Kapa Biosystems. All trademarks are the property of their respective owners. SS106001 A179 4/16 For Research Use Only. Not for use in diagnostic procedures.
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