Accel-NGS™ DNA Library Kit for Ion Torrent

DNA Library Preparation for Next Generation Sequencing (NGS) on Ion Torrent Platforms

 Catalog No  Product Name Reaction
 DL-ION1-10  Accel-NGS DNA Library Kit for Ion Torrent 10
 DL-ION1-50  Accel-NGS DNA Library Kit for Ion Torrent 50
 BC-ION1-10A  Ion Set A Barcoding Kit (10 barcodes x 1 rxn each) 10
BC-ION1-50A Ion Set A Barcoding Kit (10 barcodes x 5 rxns each) 50
 BC-ION1-10B Ion Set B Barcoding Kit (10 barcodes x 1 rxn each) 10
 BC-ION1-50B  Ion Set B Barcoding Kit (10 barcodes x 5 rxns each)  50

 

Innovative Swift technology improves sample prep for next gen sequencing by expediting the process and delivering higher quality data. The Accel-NGS DNA Library Kit for Ion Torrent platforms is the only commercially available kit capable of producing PCR-free libraries with as little as 5 ng of input DNA. PCR-free capability minimizes base composition bias and fidelity issues, while a highly efficient adapter ligation process reduces the input requirements.

  • PCR-free to minimize base composition bias and fidelity issues
  • Low input requirements: as little as 5 ng of DNA
  • Fast – only 75 minutes start-to-finish
  • Streamlined, 3-step protocol
  • Reduces adapter dimer formation to maximize sequencing output
  • Compatible with nicked, damaged, and denatured samples

Sample Types

  • Single-stranded DNA
  • Double-stranded DNA
  • Heat-denatured samples
  • Amplicons
  • Nicked DNA
  • ChIP DNA
  • FFPE DNA
  • cDNA
  • Extremely AT/GC-rich Genomes

Instrument Compatibility

  • Ion Proton System
  • Ion PGM System

Product Details

Accel-NGS technology uses a unique combination of molecular biology reagents to produce a protocol that is streamlined and fast (75 minutes start-to-finish).Two of the steps are bead-based separations that eliminate the need for time-consuming, electrophoretic gel-based size selection. Each step is color coded to make the protocol easy to follow. The simplified Accel-NGS workflow can be readily automated.

Accel-NGS DNA Library Kit for Ion Torrent Workflow

Total Turnaround Time Comparison Versus Other Kits

Note: clean-up beads are not provided with the kit. We recommend the SPRIselect Reagent Kit from Beckman Coulter.

Performance on the Ion Torrent PGM

Data below demonstrates performance of the Accel-NGS DNA Library Kit for Ion Torrent on the PGM system. Data comparing the Swift Biosciences Accel-NGS kit with other commercially available DNA library kits are also presented.

PCR-Free Libraries Prepared from 5 ng DNA

E. coli genomic DNA samples were fragmented to an average size of 260 (Figure 1a) or 180 bp (Figure 1b) using standard sonication methods. Libraries were then size-selected using SPRI beads. Accel-NGS DNA Libraries for Ion Torrent were sequenced on Ion 316 chips with either 500 (1a) or 260 (1b) flows depending on desired read length.

Figure 1. Read length histograms from 100bp and 200bp sequencing chemistries.

 

 
Sequencing Chemistry Read Length Fold Coverage Adapter Dimer  Usable Sequence Mean Accuracy
200 bp 189 bp 91.5 1.3% 56% 99.1%
100 bp 118 bp 78.0 5.3% 70% 99.6%

 

Table 1. Summary of sequencing run on 100bp and 200bp chemistries.

 

Conclusion. Accel-NGS DNA Library Kit for Ion Torrent produces sequenceable libraries PCR-free from 5 ng of input DNA. Similar performance has been obtained with input quantities ranging from 5 ng up to 8 μg.

PCR-Free versus Competitive Kit

Samples of E. coli genomic DNA (10 ng) were fragmented to an average size of 260 bp using standard sonication methods. Swift Biosciences and Company N libraries were prepared per their respective published protocols. Libraries were then size selected using SPRI beads and sequenced on Ion 316 chips with 500 flows for 200 bp chemistry.

Figure 2. Read length histograms of Swift Biosciences versus Company N.

Product PCR Cycles Read Length Fold Coverage Adapter Dimer Usable Sequence Mean Accuracy
Accel-NGS DNA Library Kit 0 182 bp 101.8 0% 61.0% 99.1%
Company N 11 152 bp 17.7 64.4% 10.6% 99.1%

 

Table 2. Summary of sequencing run of Swift Biosciences versus Company N.

Conclusion. The majority of Company N kit results were adapter dimers. Accel-NGS DNA Library Kit produces sequenceable libraries from low input without PCR, while minimizing the formation of adapter dimers to maximize sequencing output.

Great Results in About Half the Time Versus Leading Kit

1 μg samples of E. coli genomic DNA were fragmented to an average size of 260 bp using standard sonication methods. Swift Biosciences and Company L libraries were prepared per their published protocols. The Swift Biosciences library was completed in 75 minutes while Company L required 160 minutes. Both libraries were size-selected on a Pippin Prep instrument and libraries were sequenced on Ion 316 chips with 500 flows for 200 bp chemistry.

 

Figure 3. Read length histograms of Swift Biosciences versus Company L.

 

 
Product Read Length Fold Coverage Adapter Dimer  Usable Sequence Mean Accuracy
Accel-NGS DNA Library Kit 231 bp 156 0% 61% 99.4%
Company L 232 bp 118 0% 70% 99.4%

 

Table 3. Summary of sequencing run of Swift Biosciences versus Company L.

Conclusion. Accel-NGS DNA Library Kit achieves comparable performance to the leading competitive kit in about half the time.

Validation on the Ion PGM and Proton

Data below demonstrates performance of the Accel-NGS DNA Library Kit for Ion Torrent with microbial genomes of varying base composition that were prepared and sequenced on the Ion Torrent PGM and Proton systems by an independent third party. Data comparing the Accel-NGS DNA Library Kit for Ion Torrent with the leading commercially available Ion DNA library kit is also presented.

Even Coverage from 1 ng to 100 ng Input

1 ng to 100 ng of S. aureus genomic DNA (33% GC) was prepared using the Accel-NGS DNA Library Kit for Ion Torrent PCR-free protocol. Figure 1A shows the cumulative fraction of genome covered vs. depth of coverage, with a theoretical perfect result (red) when the entire genome is at normalized depth. Figure 1B shows GC content of each read vs. theoretical in silico distribution and coverage.

Figure 1A Figure 1B
 
 

Conclusion. The Accel-NGS DNA Library Kit for Ion Torrent produces libraries from an AT-rich genome with near-theoretical coverage from 100 ng down to 1 ng of input DNA.

Balanced Composition Genome Performance

 

S. pullorum genomic DNA (52% GC) was prepared using the Accel-NGS DNA Library Kit and Competitor L’s kit using their respective protocols. Similar results were obtained with E. coli, 51% GC (not shown). Figure 2A shows cumulative fraction of genome covered vs. depth of coverage, with theoretical perfect result (red) when the entire genome is at normalized depth. Figure 2B shows GC content of each read vs. theoretical in silico distribution and coverage.

Figure 2A Figure 2B
  

Conclusion. The Accel-NGS DNA Library Kit produces libraries from a balanced base composition genome with coverage comparable to Competitor L’s kit on both the PGM and Proton platforms.

 

Outperforms Leading Ion Torrent Library Prep Kit for GC-rich Genomes

 

B. pertussis genomic DNA (68% GC) was prepared using the Accel-NGS DNA Library Kit and Competitor L’s kit. Figure 3A shows cumulative fraction of genome covered vs. depth of coverage, with theoretical perfect result (red) when the entire genome is at normalized depth. Figure 3B shows GC content of each read vs. theoretical in silico distribution and coverage.

Figure 3A Figure 3B
  

Conclusion. The Accel-NGS DNA Library Kit produces libraries from a GC-rich genome with coverage closer to the theoretical perfect result vs. Competitor L’s kit on the Ion Proton.

Outperforms the Leading Ion Torrent Library Prep Kit for AT-rich Genomes

P. falciparum genomic DNA (19% GC) was prepared using the protocol from Competitor L’s kit, which required PCR . Additional genomic DNA was prepared using the Accel-NGS DNA Library Kit for Ion Torrent PCR-free protocol. Figure 4A shows the cumulative fraction of genome covered vs. depth of coverage, with the theoretical perfect result (red) when the entire genome is at normalized depth. Figure 4B shows GC content of each read vs. theoretical in silico distribution and coverage. Results demonstrated that 5% of the genome was missing in Competitor L’s library prep with PCR while < 2% of the genome was missing in Swift Biosciences library prep without PCR.

 
Figure 4A Figure 4B
  

Conclusion. The Accel-NGS DNA Library Kit produces libraries from AT-rich genomes with coverage closer to the theoretical perfect result vs. Competitor L’s kit on the Ion Proton.

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