Applications
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Preparation of DNA libraries compatible with Roche 454™ GS FLX and GS FLX Titanium™ next-generation sequencing instruments.
Nextera™ DNA Sample Prep Kits (Roche 454-Compatible) are specifically designed to prepare genomic DNA libraries compatible with Roche 454 GS FLX and GS FLX Titanium instruments. Nextera technology employs in vitro transposition* to simultaneously fragment and tag DNA in a single-tube reaction. The Nextera library preparation method offers significant advantages over existing methods, such as nebulization, sonication, or shearing (Table 1).
Benefits
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Only 50 ng of starting DNA required.
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Prepare sequencer-ready libraries in less than 2 hours.
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Incorporate platform-specific tags and optional bar codes (Fig. 1).
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Validated on Roche 454 GS FLX and GS FLX Titanium platforms.
*Covered by issued and/or pending patents.
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Fragmentation |
✓ (15-30 min) |
Add Nextera™ Enzyme Mix (5 min)
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Collection |
✓ (15 min) |
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Concentration |
✓ (15 min) |
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Size Selection |
✓ (60 min) |
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End-Repair |
✓ (60 min) |
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Clean-Up |
✓ (15 min) |
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A-Tailing |
+/- (30 min) |
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Adaptor Ligation |
✓ (60 min) |
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Clean-Up |
✓ (15 min) |
✓ (15 min) |
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Library Enrichment |
Variable (~60 min) |
PCR (~60 min) |
Blue color indicates steps where the sample is transferred to another tube.
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Roche/454-Compatible
Figure 1 (click to enlarge). Generation of libraries for Roche 454™ platforms. After fragmenting and tagging genomic DNA with Nextera™ Enzyme Mix, four-primer limited-cycle PCR is performed which adds Roche 454-compatible adaptor sequences (blue and orange). Optional bar codes (triangle) can be added between the upstream emulsion PCR (emPCR) adaptor (blue) and the transposon end (gray).
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For additional information on Nextera™ technology, please visit: www.epibio.com/nextera
For custom/bulk requests, please contact us at: Nextera@EpiBio.com
