The analysis of microbial communities by next generation sequencing, especially at low loads, can be challenging due to DNA contaminations in reagents used.
NGSeq 16S V3/V4 is a highly active, microbial DNA-free master mix that is made for the Amplicon PCR or Real-Time PCR as part of the library preparation for 16S metagenomic sequencing with Illumina® MiSeq system.
The assay contains all necessary components for the Amplicon PCR, including primers, highly active hot start Taq DNA polymerase, DNA staining solution and DNA-free water. Primers contain sequences for the amplification of the V3/V4 region of the 16S rRNA gene and the necessary overhang adaptors for Illumina. Because the assay is free of bacterial contaminations, PCRs can be run up to 40 cycles without background in negative controls.
NGSeq 16S V3/V4 is especially designed for contamination-free amplifications of bacterial DNA for best results, particularly when a deep analysis of microbial community diversification is of interest.
The discriminatory power of NGS targeting microbiomes in low load communities can be reduced not only by contaminations but by the vast number of human reads in many samples covering minor reads of bacterial sequences. Molzym offers several solutions on the ground of MolYsis™ technology and DNA-free reagents resulting in a marked increase of bacterial reads.
The combination of targeted bacterial DNA isolation and amplification with the NGSeq 16S V3/V4 allows a sample preparation without reagent-borne contaminations.