DNA Sequencing |
Next-Generation Sequencing
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Applications
- 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
- Only 50 ng of starting DNA required.
- Prepare sequencer-ready libraries in less than 2 hours.
- Incorporate platform-specific tags and optional bar codes (Fig. 1).
- Validated on Roche 454 GS FLX and GS FLX Titanium platforms.
*Covered by issued and/or pending patents.
| Fragmentation |
✓ (15-30 min) |
Add Nextera™ Enzyme Mix (5 min) |
| Collection |
✓ (15 min) |
| Concentration |
✓ (15 min) |
| Size Selection |
✓ (60 min) |
| End-Repair |
✓ (60 min) |
| Clean-Up |
✓ (15 min) |
| A-Tailing |
+/- (30 min) |
| Adaptor Ligation |
✓ (60 min) |
| Clean-Up |
✓ (15 min) |
✓ (15 min) |
| Library Enrichment |
Variable (~60 min) |
PCR (~60 min) |
Blue color indicates steps where the sample
is transferred to another tube. |
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). |
For additional information on Nextera™ technology, please visit: www.epibio.com/nextera
For custom/bulk requests, please contact us at: Nextera@EpiBio.com
Nextera™ DNA Sample Prep Kit (Illumina-Compatible)
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