Oncology NGS panels

EliGene® Adaptor-IL
  • EliGene® Adaptor-IL
  • EliGene® Adaptor-IL
  • EliGene® Adaptor-IL
  • EliGene® Adaptor-IL

EliGene® Adaptor-IL NGS

Intended use

EliGene® Adaptor-IL NGS kit is intended for the incorporation of molecular identifiers (MIDs) and specific sequencing adaptors to all amplicons generated using the multiplex PCR based on EliGene® NGS kits, enabling the identification of polymorphisms in the target area.

Principle of the method

EliGene® NGS kits enable multiplex PCR amplification of all required target regions of the gene(s) of interest. The recommended amount of DNA for each multiplex PCR reaction is between 20 and 100 ng of DNA derived from formalin-fixed paraffin-embedded (FFPE) material or from blood. Next, the resulting amplicons are barcoded, and mixed per sample. Finally, the resulting amplicon libraries of different samples are pooled and sequenced using a NGS instrument Illumina for massively parallel sequencing (MPS) according to the manufacturer’s instructions. The resulting sequence reads are subsequently analyzed by comparison with the reference sequence of the targeted gene(s) to identify variant positions.

Introduction

EliGene® NGS kits are used for one or more multiplex PCR reactions resulting in amplification of target regions of one sample. For amplification is used a ready-to-use mastermix containing a hot-start DNA polymerase. The resulting amplicons of each multiplex are diluted 1,000 fold. In the second step, a second round of PCR is performed using EliGene® Adaptor-IL NGS kit, enabling tagging of all the amplicons to incorporate MIDs and p7 and p5 adaptors required for Illumina MiSeq Sequencing. The resulting tagged amplicons from are mixed per individual applying a predefined assay specific mixing scheme. Each amplicon library is subsequently purified from small residual DNA fragments shorter than 150bp and the DNA concentration is determined using a Real-Time PCR with concentration standards. Next, these purified and individually tagged amplicon libraries are pooled equimolar, resulting in an amplicon pool or sequencing sample, which is then further processed using custom sequencing primers from EliGene® Adaptor-IL NGS kit and sequenced on the Illumina MiSeq. During the sequencing run, different sequence reads are generated on the same cluster in the process of bridge PCR. The positions of the individual custom sequencing primers supplied by EliGene® NGS kits as well as the Illumina sequencing reagents are indicated and clarified.

Specimen

Suitable specimens are the amplicons from multiplex PCR using EliGene® Lung NGS, EliGene® Colorectum NGS and EliGene® GIST NGS kits.

Ref. No. Package Min. package Price ex. VAT VAT  
EG-90069-NGS 96 reactions 1 kit € 2 000.00 21 % Buy

Instruction for Use EliGene® Adaptor-IL NGS

(published 23.01.2019, size 804.6 kB)

MSDS EliGene Adaptor-IL NGS Adaptor-IL Mix EN

(published 23.07.2018, size 528.6 kB)

MSDS EliGene Adaptor-IL NGS Index EN

(published 24.07.2018, size 528.1 kB)

MSDS EliGene Adaptor-IL NGS p501 MID Primer EN

(published 23.07.2018, size 528.3 kB)

MSDS EliGene Adaptor-IL NGS p502 MID Primer EN

(published 23.07.2018, size 528.5 kB)

MSDS EliGene Adaptor-IL NGS p503 MID Primer EN

(published 23.07.2018, size 528.5 kB)

MSDS EliGene Adaptor-IL NGS p504 MID Primer EN

(published 23.07.2018, size 528.5 kB)

MSDS EliGene Adaptor-IL NGS p505 MID Primer EN

(published 23.07.2018, size 534.9 kB)

MSDS EliGene Adaptor-IL NGS p506 MID Primer EN

(published 23.07.2018, size 528.4 kB)

MSDS EliGene Adaptor-IL NGS p701 MID Primer EN

(published 23.07.2018, size 535.3 kB)

MSDS EliGene Adaptor-IL NGS p702 MID Primer EN

(published 23.07.2018, size 528.6 kB)

MSDS EliGene Adaptor-IL NGS p703 MID Primer EN

(published 23.07.2018, size 535.2 kB)

MSDS EliGene Adaptor-IL NGS p704 MID Primer EN

(published 23.07.2018, size 528.5 kB)

MSDS EliGene Adaptor-IL NGS p705 MID Primer EN

(published 23.07.2018, size 528.6 kB)

MSDS EliGene Adaptor-IL NGS p706 MID Primer EN

(published 23.07.2018, size 535.3 kB)

MSDS EliGene Adaptor-IL NGS p707 MID Primer EN

(published 23.07.2018, size 528.6 kB)

MSDS EliGene Adaptor-IL NGS p708 MID Primer EN

(published 23.07.2018, size 528.5 kB)

MSDS EliGene Adaptor-IL NGS RP1 Primer EN

(published 23.07.2018, size 528.1 kB)

MSDS EliGene Adaptor-IL NGS RP2 Primer EN

(published 23.07.2018, size 528.1 kB)

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