NucleoMag NGS Clean-up and Size Selection für NGS Applikationen

655,00 €*

Packungseinheit 50 mL

*ohne Steuern und Versandkosten

REF 744970.50

Packungseinheit 50 mL

Verkaufseinheit

5 mL

50 mL

500 mL

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Das NucleoMag NGS Clean-up and Size Select Kit ermöglicht die maßgeschneiderte Vorbereitung von NGS-Bibliotheken mit einfachen Clean-up sowie ein- oder beidseitigen Größenauswahlprotokollen. Das Kit ermöglicht eine präzise Feinabstimmung der Größenselektion mit einer Genauigkeit von 50 bp. Es ist möglich, DNA mit einem gewählten Größenschwellenwert oder innerhalb eines bestimmten Größenbereichs zu selektieren. Die Beads eignen sich auch für die einfache Reinigung von Nukleinsäuren aus Reaktionsgemischen, z. B. bei der cDNA-Synthese oder bei In-vitro-Transkriptionsreaktionen.

Anwendung	Clean-up und Größenauswahl für NGS-Bibliotheksvorbereitungen
Automatische Nutzung	Ja
CE-zertifiziert	Nein, nur für wissenschaftlichen Gebrauch
Elutionsvolumen	10−100 µL
Format	Magnetic beads
Fragmentgröße	150 bp−ca. 800 bp
Gefahrstoffe	Nein
Haltbarkeit (ab Produktion)	24 Monat(e)
Handhabung	Magnetische Trennung
Lagertemperatur	2–8 °C
Präparationszeit	40−120 min/96 Präps
Probenmaterial	Reaktionsgemische aus NGS-Bibliothekskits
Probenmenge	7,5 pg-5 µg Nukleinsäuren in NGS-Reaktionsmischungen
Technologie	Magnetische Bead-Technologie
Typische Downstream-Anwendung	Next Generation Sequenzierung
Typische Rückgewinnung	≥ 80 %
Verkaufseinheit	5 mL, 50 mL, 500 mL
Warenzeichen	NucleoMag
Ziel	Clean up

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Instruction NucleoMag NGS Clean-up and Size Select

Flyer NucleoMag NGS Clean-up and Size Select

NucleoMag NGS Clean-up and Size Select on Opentrons OT-2

Instruction NucleoMag NGS Clean-up and Size Select (FR)

Leaflet

Magnetic bead-based DNA and RNA Clean-up

NucleoMag NGS Clean-up and Size Select

Most sequencing platforms for library preparations require enzymatic reaction Clean-up and fragment size selection. NucleoMag NGS Clean-up and Size Select enables Clean-up reactions, as well as single or double sided size selection, to recover the fragment lengths which are needed in your specific application. Simply dilute the magnetic beads to utilize the tunable size selection feature. Dilution ratios are similar to most other comparable magnetic bead products in the market, allowing for the NucleoMag NGS Clean-up and Size Select kit to be used with your current protocols

Product at a glance

Technology	Magnetic beads
Applications	Size selection for next-generation-sequencing workflows PCR Clean-up RNA or cDNA Clean-up after purifications or enzymatic reactions (i. e. IVT or DNA digestion)
Recovery	70 – 100 %
Elution volume	10 – 100 μL
Shelf-life (from production)	24 months
Product sizes	5 mL, 50 mL, 500 mL

Schematic Workflow Overview

1. Add bead suspension to your DNA or RNA solution

2. Incubation of magnetic beads with sample for proper binding

3. Removal of contaminants by ethanol washes

4. Dry magnetic beads to allow evaporation of ethanol traces

5. Elution of DNA or RNA

6. Transfer of the final product to a new plate/tube

One Clean-up kit for all samples!

Use your current protocol. No changes needed.

Application data – DNA Clean-Up and Size Selection

Size selection of fragment mix

For single sided size selection (left or right), the sample is mixed with the beads in predetermined ratios for the desired exclusion of smaller or larger sized fragments. For the double sided size selection, two binding steps are performed to exclude larger fragments above the cut-off and smaller fragments below the lower cut-off.

Reliable reproducibility in DNA clean-up procedures

To assess reproducibility of DNA clean-up, EcoRI-linearized pcDNA 3.1(+) and Ndel-linearized pGEM inserts were purified with four replicates each. For DNA purification, a sample to NGS clean-up suspension-ratio of 1.0 was used. The parallel Clean-up experiments show high reproducibility and recovery of the target DNA.

Fragment size analysis of prepared NGS libraries

NGS libraries were prepared using Truseq* DNA PCR Free kit from Illumina (red), AMPure* XP (blue) and NucleoMag NGS Clean-up and Size Select (green) for Clean-up and size selection steps. (A) Input DNA, 1 μg sheared E. coli DNA. (B) Size distribution of DNA Fragments after library preparation as input for sequencing with expected fragment size of 650 bp (insert + adapters). Data kindly provided by TAKARA BIO INC.

Sequencing analysis

In order to compare the sequencing performance after library preparation generation different clean-up and size selection products were tested. Performed tests included the distribution of genome sequence coverage (depth of genome base coverage) and effect on GC content on sequence coverage (A) No significant difference in mapped depth of genome base coverage. The constant depth indicates low bias. (B) Horizontal plot pattern of MN beads compared to AMPure* XP beads. After the summary of aligned read bases within a 0.5 kb genome region, the two samples have been normalized / matched resulting in a horizontally pattern showing the similarity of the two samples. Data kindly provided by TAKARA BIO INC.

Cited in more than 50 publications

The NucleoMag NGS Clean-up and Size Select has been cited in more than 50 peer-reviewed publications. The kit has been used for PCR Clean-ups as well as in next generation sequencing workflows.

More Information

Application data – RNA Clean-Up

RNA Clean-Up with reliable recovery rates as well as high purities

A) MACHEREY‑NAGEL’s NucleoMag NGS Clean-up and Size Select beads as well as RNAClean* XP beads from Beckman Coulter (BC) were used for Clean-up of RiboRuler High Range Ladder, resulting in recovery rates of 88.6 ± 7.4 % (MN) and 81.5 ± 5.4 % (BC). B) RNA isolated from HeLa cells were used as input for Clean-up reactions which generated improved purities.

High-integrity RNA after Clean-up with NucleoMag NGS Clean-Up and Size Select kit

RNA samples were cleaned-up using the NucleoMag NGS Clean-up and Size Select kit (MACHEREY‑NAGEL) as well as RNAClean* XP beads (Beckman Coulter). 500 ng RNA was used as starting material. The RNA was eluted in 25 μL and analyzed on Agilent’s Bioanalyzer 2100. No significant difference in RNA integrity has been observed for used products, as integrity values after clean-up are comparable to the input sample (RIN ≥ 9,4).

Product	ng/μL	A260 / A280	A260 / A230
MN Beads	14.4 ± 2.1	2.3 ± 0.1	2.5 ± 0.7
RNAClean* XP	14.4 ± 1.6	2.3 ± 0.1	2.2 ± 0.8

NucleoMag NGS Clean-Up and Size Select kit is suitable for Clean-ups of RNA transcripts after in vitro transcription reactions

Riboprobe* System – T7 has been used for in vitro transcription of single-stranded RNA using a pGEM* vector. NucleoMag NGS Clean-up and Size Select and RNAClean* XP were used for Clean-up of IVT reactions.