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PGN001  PureGenome™ Next Generation Sequencing Library DNA Modifier

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24 May 2013

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The PureGenome™ High Efficiency Next Generation Sequencing (NGS) Library Preparation Reagents are uniquely optimized as a one-stop solution for NGS Library Preparation. Validated for sequencing compatibility with Illumina® instruments; three sets of reagents are offered each having a unique functionality in the library preparation work flow.

The PureGenome NGS Library DNA Modifier, when purchased with the PureGenome NGS Library A-Tail Enhancer, includes all of the components needed for end repair, dA-tailing, and adapter ligation of user supplied sheared input DNA. The enzymes and buffers are supplied in easy-to-use formulations. These reagents, when used together, provide a streamlined, two-step solution for preparing adapter-ligated DNA libraries for NGS (See Figure 1 at bottom of this page.).

The PureGenome NGS Library Amplifier contains the ultra-high fidelity and fast yielding KOD Hot Start DNA Polymerase for minimal bias PCR NGS library enrichment. KOD Hot Start DNA Polymerase allows high yielding amplification of challenging DNA templates (high GC or TA repeat sequence content). Collectively, this solution offers rapid amplification of NGS libraries for ensuring required inputs for NGS instrument sequencing and high quality data output.

The PureGenome High Efficiency Next Generation Sequencing (NGS) Library Preparation reagents sets, purchased as one NGS library preparation solution, are formulated to allow construction of 50 NGS Libraries when following the protocol as found in Technical Bulletin 574 (TB574).

*Note: PGN001-1EA, PGN002-1EA, PGN003-1EA should be purchased as a set for a lot-matched quality controlled solution for NGS library preparation.


Components


• 1 EA          PureGenome NGS Library DNA Modifier (PGN001-1EA)
  • 0.5 ml      PureGenome Conversion Buffer
  • 0.5 ml      PureGenome 10 mM ATP
  • 0.2 ml      PureGenome 10 mM dNTP mix
  • 750 units   PureGenome T4 DNA polymerase
  • 2500 units  PureGenome T4 Polynucleotide Kinase
  • 0.5 ml      PureGenome Ligation Buffer
  • 0.5 ml      PureGenome DNA Ligase
• 1 EA          PureGenome NGS Library A-tail Enhancer (PGN002-1EA)
  • 2.4 ml      PureGenome NovaTaq™ DNA Polymerase
• 1 EA          PureGenome NGS Library Amplifier (PGN003-1EA)
  • 1.25 ml     KOD Hot Start Master Mix

Storage: Store all components at –20 °C. All reagents are stable for at least one year at the recommended storage temperature.


Representative Data

See Figure 2 at the bottom of this page for representative data.


Store and ship information
Storage -20°C
Ship Blue Ice or Dry Ice
Standard Handling
Data

Figure 1: PureGenome High Efficiency NGS Library Preparation Workflow

Figure 2: Maximim coverage with minimum bias

© Merck KGaA, Darmstadt, Germany, 2013

All references to Merck refer to Merck KGaA, Darmstadt, Germany.


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