NucleoMag NGS Clean-up and Size Select

NucleoMag NGS Clean-up and Size Select employs scalable, automation-friendly magnetic bead technology to enable efficient cleanup of enzymatic reactions and tunable size selection of DNA fragments generated in NGS library preparation workflows. The flexible protocol enables processing of 50–150 µl input volumes and allows for enrichment of fragments ranging in size from 150–800 bp. NucleoMag NGS Clean-up and Size Select consists of paramagnetic beads suspended in binding buffer that selectively bind DNA fragments based on the volume ratio of bead suspension to sample. After magnetic separation and removal of supernatant, the beads are washed with ethanol followed by a short drying step. Following elution in low-salt buffer or water, the highly purified DNA is free of contaminants such as nucleotides, primers, adapters, adapter dimers, enzymes, buffer additives, or salts, and can be used directly in downstream applications. NucleoMag NGS Clean-up and Size Select provides comparable performance and employs the same dilution ratios for size selection as magnetic bead-based solutions from other providers, such that it can be incorporated in existing workflows without the need for protocol adjustment or optimization.

Cat. # 744970.5 consists of a bottle containing 5 ml of bead suspension.

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Tunable selection of DNA fragments in a given size range using NucleoMag NGS Clean-up and Size Select

Tunable selection of DNA fragments in a given size range using NucleoMag NGS Clean-up and Size Select. NucleoMag NGS Clean-up and Size Select can be used to isolate DNA fragments of a given size within a target range of ~150�800 bp. Selective binding of DNA fragments above a designated size cutoff is achieved by applying NucleoMag beads to the sample at a predetermined dilution ratio. This approach can be applied once or twice to perform single- or double-sided size selection. In the latter case, an initial round of bead purification is typically performed to bind and remove fragments above the upper size cutoff, and a second round of purification is performed to bind and retain fragments above the lower size cutoff. Dilution ratios used for size selection with NucleoMag beads are similar to ratios employed by size-selection beads from other providers, such that NucleoMag beads can be incorporated in existing NGS library preparation workflows without protocol adjustment.

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Relationship between bead-to-sample volume ratio and recovery rate for DNA fragments of different sizes with NucleoMag NGS Clean-up and Size Select

Relationship between bead-to-sample volume ratio and recovery rate for DNA fragments of different sizes with NucleoMag NGS Clean-up and Size Select. Varying quantities of NucleoMag beads were applied to 100-?l aliquots containing mixtures of DNA fragments ranging in size from 100 bp to 1,000 bp (10 ng/?l) in the indicated dilution ratios (gray row). Recovery rates for fragments of each size are expressed as a percentages (%).

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Examples of double-sided size selection using NucleoMag NGS Clean-up and Size Select

Examples of double-sided size selection using NucleoMag NGS Clean-up and Size Select. Sheared mouse genomic DNA was size selected with NucleoMag beads using two different pairs of dilution ratios and measured using a Bioanalyzer. Green trace: sheared mouse genomic DNA prior to size selection. Red trace: size distribution after double-sided size selection using dilution ratios of 0.4 (right) and 0.6 (left), resulting in a mean fragment size of 460 bp. Blue trace: size distribution after double-sided size selection using dilution ratios of 0.55 (right) and 0.8 (left), resulting in a mean fragment size of 340 bp.

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Fragment size analysis for NGS libraries prepared using size-selection beads from various providers

Fragment size analysis for NGS libraries prepared using size-selection beads from various providers. NGS libraries were prepared from sheared E. coli DNA using the TruSeq DNA PCR-Free kit by Illumina and size selected using magnetic beads from different providers (Illumina, AMPure XP, and NucleoMag NGS Clean-up and Size Select). Panel A. Bioanalyzer trace indicating size distribution of sheared E. coli DNA (1-?g input) prior to NGS library preparation. Panel B. Bioanalyzer traces indicating size distributions of resulting NGS libraries purified using Illumina (red), AMPure XP (blue), or NucleoMag (green) magnetic beads.

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Comparison of sequencing data generated from NGS libraries prepared using size-selection beads from various providers

NGS libraries were prepared from sheared E. coli DNA using the TruSeq DNA PCR-Free kit by Illumina and size selected using magnetic beads from different providers (Illumina, AMPure XP, and NucleoMag NGS Clean-up and Size Select). Panel A. Bioanalyzer trace indicating size distribution of sheared E. coli DNA (1-?g input) prior to NGS library preparation. Panel B. Bioanalyzer traces indicating size distributions of resulting NGS libraries purified using Illumina (red), AMPure XP (blue), or NucleoMag (green) magnetic beads.

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744970.5: NucleoMag NGS Clean-up and Size Select

NucleoMag NGS Clean-up and Size Select employs scalable, automation-friendly magnetic bead technology to enable efficient cleanup of enzymatic reactions and tunable size selection of DNA fragments generated in NGS library preparation workflows. The flexible protocol enables processing of 50–150 µl input volumes and allows for enrichment of fragments ranging in size from 150–800 bp. NucleoMag NGS Clean-up and Size Select consists of paramagnetic beads suspended in binding buffer that selectively bind DNA fragments based on the volume ratio of bead suspension to sample. After magnetic separation and removal of supernatant, the beads are washed with ethanol followed by a short drying step. Following elution in low-salt buffer or water, the highly purified DNA is free of contaminants such as nucleotides, primers, adapters, adapter dimers, enzymes, buffer additives, or salts, and can be used directly in downstream applications. NucleoMag NGS Clean-up and Size Select provides comparable performance and employs the same dilution ratios for size selection as magnetic bead-based solutions from other providers, such that it can be incorporated in existing workflows without the need for protocol adjustment or optimization.

Cat. # 744970.50 consists of a bottle containing 50 ml of bead suspension.

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Tunable selection of DNA fragments in a given size range using NucleoMag NGS Clean-up and Size Select

Tunable selection of DNA fragments in a given size range using NucleoMag NGS Clean-up and Size Select. NucleoMag NGS Clean-up and Size Select can be used to isolate DNA fragments of a given size within a target range of ~150�800 bp. Selective binding of DNA fragments above a designated size cutoff is achieved by applying NucleoMag beads to the sample at a predetermined dilution ratio. This approach can be applied once or twice to perform single- or double-sided size selection. In the latter case, an initial round of bead purification is typically performed to bind and remove fragments above the upper size cutoff, and a second round of purification is performed to bind and retain fragments above the lower size cutoff. Dilution ratios used for size selection with NucleoMag beads are similar to ratios employed by size-selection beads from other providers, such that NucleoMag beads can be incorporated in existing NGS library preparation workflows without protocol adjustment.

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Examples of double-sided size selection using NucleoMag NGS Clean-up and Size Select

Examples of double-sided size selection using NucleoMag NGS Clean-up and Size Select. Sheared mouse genomic DNA was size selected with NucleoMag beads using two different pairs of dilution ratios and measured using a Bioanalyzer. Green trace: sheared mouse genomic DNA prior to size selection. Red trace: size distribution after double-sided size selection using dilution ratios of 0.4 (right) and 0.6 (left), resulting in a mean fragment size of 460 bp. Blue trace: size distribution after double-sided size selection using dilution ratios of 0.55 (right) and 0.8 (left), resulting in a mean fragment size of 340 bp.

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Comparison of sequencing data generated from NGS libraries prepared using size-selection beads from various providers

NGS libraries were prepared from sheared E. coli DNA using the TruSeq DNA PCR-Free kit by Illumina and size selected using magnetic beads from different providers (Illumina, AMPure XP, and NucleoMag NGS Clean-up and Size Select). Panel A. Bioanalyzer trace indicating size distribution of sheared E. coli DNA (1-?g input) prior to NGS library preparation. Panel B. Bioanalyzer traces indicating size distributions of resulting NGS libraries purified using Illumina (red), AMPure XP (blue), or NucleoMag (green) magnetic beads.

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744970.50: NucleoMag NGS Clean-up and Size Select

NucleoMag NGS Clean-up and Size Select employs scalable, automation-friendly magnetic bead technology to enable efficient cleanup of enzymatic reactions and tunable size selection of DNA fragments generated in NGS library preparation workflows. The flexible protocol enables processing of 50–150 µl input volumes and allows for enrichment of fragments ranging in size from 150–800 bp. NucleoMag NGS Clean-up and Size Select consists of paramagnetic beads suspended in binding buffer that selectively bind DNA fragments based on the volume ratio of bead suspension to sample. After magnetic separation and removal of supernatant, the beads are washed with ethanol followed by a short drying step. Following elution in low-salt buffer or water, the highly purified DNA is free of contaminants such as nucleotides, primers, adapters, adapter dimers, enzymes, buffer additives, or salts, and can be used directly in downstream applications. NucleoMag NGS Clean-up and Size Select provides comparable performance and employs the same dilution ratios for size selection as magnetic bead-based solutions from other providers, such that it can be incorporated in existing workflows without the need for protocol adjustment or optimization.

Cat. # 744970.500 consists of a bottle containing 500 ml of bead suspension.

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Relationship between bead-to-sample volume ratio and recovery rate for DNA fragments of different sizes with NucleoMag NGS Clean-up and Size Select

Relationship between bead-to-sample volume ratio and recovery rate for DNA fragments of different sizes with NucleoMag NGS Clean-up and Size Select. Varying quantities of NucleoMag beads were applied to 100-?l aliquots containing mixtures of DNA fragments ranging in size from 100 bp to 1,000 bp (10 ng/?l) in the indicated dilution ratios (gray row). Recovery rates for fragments of each size are expressed as a percentages (%).

Back

Fragment size analysis for NGS libraries prepared using size-selection beads from various providers

Fragment size analysis for NGS libraries prepared using size-selection beads from various providers. NGS libraries were prepared from sheared E. coli DNA using the TruSeq DNA PCR-Free kit by Illumina and size selected using magnetic beads from different providers (Illumina, AMPure XP, and NucleoMag NGS Clean-up and Size Select). Panel A. Bioanalyzer trace indicating size distribution of sheared E. coli DNA (1-?g input) prior to NGS library preparation. Panel B. Bioanalyzer traces indicating size distributions of resulting NGS libraries purified using Illumina (red), AMPure XP (blue), or NucleoMag (green) magnetic beads.

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744970.500: NucleoMag NGS Clean-up and Size Select