: New Technology for PCR-Free & Low-Input NGS Libraries
TM
Laurie Kurihara, Ron Beaubien, Jeff Perry, Vanessa Kelchner, Drew McUsic,
Rachel Spurbeck, Julie Laliberte, Sergey Chupreta and Vladimir Makarov
Swift Biosciences, Inc. 58 Parkland Plaza, Suite 100 Ann Arbor, MI 48103 734.330.2568
TM
Abstract
Accel-NGS Excels in Turn Around Time
Next-generation sequencing (NGS) has accelerated genomic studies across a wide
range of sample types. Library construction from these samples for whole
genome, exome or targeted gene panel sequencing is characterized by long
preparation times and sample constraints due to DNA input quantity and
molecular integrity. Accel-NGS is a novel technology that enables faster and more
efficient library preparation. Accel-NGS libraries can be produced from DNA input
quantities as low as 5 ng without the need for PCR amplification, thereby
minimizing base composition bias. The streamlined workflow requires 75 minutes
start-to-finish and has only 5 steps, making it easily automatable. Accel-NGS is
also compatible with damaged or cross-linked samples including FFPE because it
does not require intact, double-stranded DNA for adapter ligation. When
compared with commercially available kits, Accel-NGS libraries sequenced on the
Ion Torrent PGM™ demonstrated higher library yield and lower adapter dimer
formation, thus maximizing sequence data output from low-input DNA quantity.
Company N
Company L
Accel-NGS
0
50
100
150
200
Total Time (min.)
Great Results in About Half the Time
Accel-NGS
70000
60000
60000
50000
50000
Count
Count
Accelerate Your NGS Library Prep
40000
30000
40000
30000
20000
20000
10000
10000
0
0
50
100
150
200
Company L
70000
250
300
350
50
100
Read Length
• Only kit on the market for low input without PCR
• No PCR - minimizes base composition bias or fidelity issues
• Unique two-step adaptation process reduces adapter dimers and nonfunctional molecules
250
300
350
Read Length
Accel-NGS 231 bp
Company L 232 bp
• Fast: Only 75 minutes start-to-finish
200
Read
Fold
Adapter
Usable
Mean
Length Coverage Dimer Sequence Accuracy
Product
• Highly efficient adaptation allows for ultra-low input quantities (5 ng)
150
156
118
0%
0%
61%
70%
99.4%
99.4%
• 1 mg samples of E. coli genomic DNA were fragmented to an average size of 260 bp
• Company L libraries were prepared per their published protocol
• Libraries were size selected on a Pippin Prep™
• Compatible with FFPE and damaged samples
• Libraries were sequenced on Ion 316 chips with 500 flows for 200 bp chemistry
• No need for time-consuming gel-based size selection
Accel-NGS Excels with Ultra -Low Input Levels
Accel-NGS: Libraries from 5 ng Without PCR Amplification
Accel-NGS
20000
18000
20000
Count
90000
18000
80000
16000
70000
14000
12000
Count
Count
100 bp Read
100000
10000
8000
60000
50000
40000
6000
30000
4000
20000
2000
18000
16000
16000
14000
14000
12000
12000
Count
200 bp Read
10000
8000
10000
8000
6000
6000
4000
4000
2000
2000
0
0
50
100
150
200
Company N
20000
250
300
50
350
100
150
200
250
300
350
Read Length
Read Length
10000
0
50
100
150
200
250
300
0
350
50
Read Length
100
150
200
250
300
350
Read Length
Read
Fold
Adapter Usable
Mean
Sequencing Chemistry
Length Coverage Dimer Sequence Accuracy
200 bp
100 bp
189 bp
118 bp
91.5
78.0
1.3%
5.3%
56%
70%
99.1%
99.6%
• 5 ng samples of E. coli genomic DNA were fragmented to an average size of 260 (left) or
180 bp (right)
• Libraries were size selected using SPRI® beads
• Accel-NGS Libraries were sequenced on Ion 316™ chips with either 500 (left) or 260 (right)
flows depending on desired read length
• Similar performance is observed for input quantities ranging from 5 ng to 8 mg
Accel-NGS is for Research Use Only
Ion Torrent, Ion 316, and PGM are registered trademarks of Ion Torrent Systems, Inc., now part of Life Technologies Corp.
Pippin Prep is a registered trademark of Sage Science, Inc.
SPRI is a registered trademark of Beckman Coulter, Inc.
Read length histograms and alignments to the reference genome were generated using Torrent Suite 3.0 software
Product
Accel-NGS
Company N
Company L*
PCR
Read
Fold
Adapter
Usable
Mean
Cycles Length Coverage Dimer Sequence Accuracy
0
11
-
182 bp
152 bp
-
101.8
17.7
-
0%
64.4%
-
61.0%
10.6%
-
99.1%
99.1%
-
• Majority of Company N kit results were adapter dimers
• 10 ng samples of E. coli genomic DNA were fragmented to an average size of 260 bp
• Company N libraries were prepared per their published protocol
• Libraries were size selected using SPRI beads
• Libraries were sequenced on Ion 316 chips with 500 flows for 200 bp chemistry
*Company L could not be included in this comparison because the vendor's protocol requires > 50 ng input DNA
Accel-NGS Library Prep Technology
• PCR-free libraries from as low as 5 ng input DNA
• Fast: Only 75 minutes start-to-finish
• Prevents formation of adapter dimers to maximize sequencing output
• Compatible with FFPE and damaged samples
www.swiftbiosci.com