Oncology Test Kit

EliGene® NGS kits facilitate the multiplex PCR amplification of all necessary target regions of the gene(s) of interest. Between 20 and 100 ng of DNA derived from formalin-fixed paraffin-embedded (FFPE) material or from blood is the recommended amount of DNA for each multiplex PCR reaction. The resulting amplicons are subsequently mixed per sample and barcoded with MID sequences. The manufacturer's instructions are followed to quantify and sequence this sample using an Illumina NGS instrument for massively parallel sequencing (MPS). Subsequently, the sequence reads are compared to the reference sequence of the targeted gene(s) to identify variant positions.input:
output:Opening Statementinput:
output:amplification of target regions of a single sample is achieved through the use of EliGene® NGS kits in one or more multiplex PCR reactions. A hot-start DNA polymerase is included in a ready-to-use mastermix for amplification. The amplicons of each multiplex that are produced are diluted by a factor of 100. A second round of PCR is conducted in the second step, utilizing the EliGene® Adaptor-IL NGS kit to tag all amplicons with MIDs and p7 and p5 adaptors that are necessary for Illumina MiSeq Sequencing. The tagged amplicons that are produced are mixed per individual using a predetermined assay-specific mixing scheme. The DNA concentration is determined using a Real-Time PCR with concentration standards, and each amplicon library is subsequently purified from small residual DNA fragments shorter than 150bp.