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Applications
- Preparation of DNA libraries compatible with Illumina Solexa® next-generation
sequencing instruments.
The Nextera™ DNA Sample Prep Kit (Illumina-Compatible) is specifically
designed to prepare genomic DNA libraries compatible with Illumina Solexa GAI
and GAII 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 Illumina Solexa GAI and GAII instruments.
*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. |
Illumina/Solexa-Compatible
Figure 1 (click to enlarge). Generation of libraries for Illumina
Solexa® platforms. Target DNA is fragmented and tagged with
Nextera™ Enzyme Mix containing ends appended with sequencing
primer sites (blue and orange). Limited-cycle PCR with a four-primer
reaction adds bridge PCR (bPCR) compatible adaptors (purple and pink)
to the core sequencing library. Optional bar codes (triangle) can be
added between the downstream bPCR adaptor (pink) and the core sequencing
library adaptor (orange). Alternative sequencing primers are required
for the Illumina Solexa-compatible libraries: Read 1 Primer (blue/gray
arrow); Read 2 Primer (orange/gray arrow); and Index Read Primer (gray/orange
arrow). |
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 Kits (Roche 454-Compatible)
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