Nextera™ DNA Sample Prep Kit (Illumina-Compatible)
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 vitrotransposition* 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).
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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
Nextera™ DNA Sample Prep Kits (Roche 454-Compatible)
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
