二代测序NGS样品制备试剂盒

NGS二代测序样品制备试剂盒

2012年下架，生产线调整，推出新产品，隶属Illumina公司

GA09115	Nextera™ DNA Sample Prep Kit (Illumina-compatible)	5 Rxns
GA091120	Nextera™ DNA Sample Prep Kit (Illumina-compatible)	20 Rxns
GA0911-50	Nextera™ DNA Sample Prep Kit (Illumina-compatible)	50 Rxns
GA0911-96	Nextera™ DNA Sample Prep Kit (Illumina-compatible)	96 Rxns
GABC0950	Nextera™ Bar Codes (Illumina-compatible)	12 Bar Codes
NT09115	Nextera™ DNA Sample Prep Kit (Roche Titanium-compatible)	5 Rxns
NT091120	Nextera™ DNA Sample Prep Kit (Roche Titanium-compatible)	20 Rxns
NT0911-50	Nextera™ DNA Sample Prep Kit (Roche Titanium-compatible)	50 Rxns
NT0911-96	Nextera™ DNA Sample Prep Kit (Roche Titanium-compatible)	96 Rxns
NTBC0950	Nextera™ Bar Codes (Roche Titanium-compatible)	12 Bar Codes
FL09115	Nextera™ DNA Sample Prep Kit (Roche FLX-compatible)	5 Rxns
FL091120	Nextera™ DNA Sample Prep Kit (Roche FLX-compatible)	20 Rxns
FLBC0950	Nextera™ DNA Sample Prep Kit (Roche FLX-compatible)	50 Rxns
FLBC0950	Nextera™ Bar Codes (Roche FLX-compatible)	12 Bar Codes
EM091120	Nextera™ PCR Enzyme	20 ul
EM091150	Nextera™ PCR Enzyme	50 ul
EM0911-96	Nextera™ PCR Enzyme	96 ul

Nextera™ Technology for Next-Generation Sequencing Library Preparation EPICENTRE's revolutionary Nextera™ technology* uses in vitro transposition to prepare sequencer-ready libraries from genomic DNA for multiple sequencing platforms. The technology simultaneously fragments and tags DNA, in a single-tube reaction. Use nanogram amounts of starting DNA. Prepare sequencer-ready libraries in less than 2 hours. Incorporate platform-specific tags and optional barcodes. Validated on Roche 454™ GS FLX and GS FLX Titanium™, and Illumina® Solexa® GAI, GAII. Library Preparation Comparison Processing Step Standard (~µg) Nextera™ (50 ng) 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) Total Time: ~6 Hours <2 Hours

Nextera™ Technology Overview

Nextera technology employs in vitro transposition to prepare sequencer-ready libraries. In a modification of the classic transposition reaction, we used free transposon ends and a transposase to form a Transposome™ complex. When this complex is incubated with target double-stranded DNA (dsDNA), the target is fragmented and the transferred strand of the transposon end oligonucleotide is covalently attached to the 5´ end of the target fragment (Fig. 1).

Figure 1. Simultaneous DNA fragmentation and tagging by in vitro cut-and-paste transposition. When free transposon ends are used in the insertion reaction, the target DNA is cleaved and tagged at the 5´ end with the transposon sequence. The resulting fragments have single-stranded gaps.

Nextera technology can be used to generate di-tagged libraries, with optional barcoding, compatible with Roche/454 or Illumina/Solexa® platforms. To generate platform-specific libraries, either free transposon ends (Fig. 2A) or appended transposon ends (Fig. 2B) can be used. Platform-specific tags (and optional bar coding) are introduced by 10 cycles of PCR. The sequencing adaptors enable amplification by emulsion PCR (emPCR), bridge PCR (bPCR), and other methods. The amplified library can be subsequently sequenced using the appropriate primers.

A. Roche/454-Compatible	Figure 2. Generation of platform-specific sequencing libraries. A) Target DNA is fragmented and tagged with the Nextera™ Enzyme Mix. Limited-cycle PCR with a four-primer reaction adds Roche/454-compatible adaptors sequences (blue and orange). Optional bar coding (triangle) is added between the upstream emPCR adaptor (blue) and the transposon end (gray). B) Target DNA is fragmented and tagged with the Nextera Enzyme Mix containing ends appended with sequencing primer sites (blue and orange). Limited-cycle PCR with a four-primer reaction adds bPCR-compatible adaptors (purple and pink) to the core sequencing library. Optional bar coding (triangle) is 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 (blue/gray arrow); Read 2 (orange/gray arrow); Index Read (gray/orange arrow).
B. Illumina/Solexa-Compatible