Features and Benefits

Norgen's NGS Normalization 96-Well Kit allows for high-throughput NGS library PCR amplicon clean-up and normalization of NGS library PCR product concentration. The clean-up removes all PCR by-products including primers, dimers, enzymes and unincorporated nucleotides during the library prep PCR step. Purification is based on 96-well column chromatography using Norgen’s proprietary resin as the separation matrix. The amount of Norgen’s matrix in each 96 well is optimized to have a limited binding capacity, thus each well can elute an equal concentration of PCR product. The process involves first adding 3 volumes of Buffer SK to the PCR product, and the mixture is then loaded onto the 96-Well Normalization Plate. The bound PCR amplicon is then washed with the provided Wash Solution A in order to remove any remaining impurities, and the purified PCR amplicon is eluted with Elution Buffer B. The purified and normalized library PCR amplicon can then be used in NGS workflows and other sequencing applications.

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Supporting Data

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Figure 1. The V4 region of the 16S rRNA gene (~250bp) was amplified in 20 µL of PCR reaction using 16s indexed primers and normalized using Norgen’s NGS Normalization 96-Well Kit. The kit successfully removes primer dimers and normalizes the concentration in the 100 µL of elution. A: Before Normalization (Original PCR product), B: After normalization with Norgen’s NGS normalization.

Figure 2. Comparison with SequalPrep Normalization Plate (96) Kit (Invitrogen) for the normalized DNA yield. Fifty PCR products (20 µL reaction) from the same PCR reaction batch were randomly divided and normalized using the two different kits. The larger elution volume and higher concentration provided with Norgen's NGS Normalization Kit allows for repeated and future assays to be performed.

Figure 3. The efficiency of the DNA normalization in comparison with SequalPrep (Invitrogen). Norgen's NGS Normalization 96-Well Kit showed a lower error percentage compared to Invitrogen's SequalPrep Kit, indicating the uniformed normalization performance of Norgen's kit.

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