Reagent Kits

AATI's capillary electrophoresis instruments utilize specially designed reagent kits to analyze diverse nucleic acid samples, from small RNA to genomic DNA, and NGS library quality control.

Reagent Kit Box

FP-1101-0275

Ultra-Sensitivity NGS Analysis Kit

AccuCleave Agarose Kits
Ultra-Sensitivity NGS Analysis Kit

The Ultra-Sensitivity NGS Analysis Kit for the FEMTO Pulse™ Automated Pulsed-Field CE Instrument provides unprecedented detection sensitivity. DNA fragments and smears are precisely quantified and qualified into the femtogram range. Conserve precious sample and quantify and qualify NGS libraries for challenging applications including PCR-free libraries and single cell genomics.

Components

275 Sample Kit
NGS Separation 5101 Gel 250 mL (FP-5101-0250)
FEMTO Pulse Intercalating Dye 30 µL (FP-6001-U030)
5X Inlet Buffer 75 mL (DNF-325-0075)
5X Conditioning Solution 50 mL (DNF-425-0050)
Ultra-Sensitivity NGS Diluent Marker 25 µL (FP-8101-U025)
Ultra-Sensitivity NGS Ladder 60 µL (FP-7101-U060)
Dilution Buffer 0.25X TE 12 mL (DNF-498-0012)
0.25X TE Rinse Buffer 60 mL (DNF-497-0060)
BF-P25 Blank Solution 5 mL (DNF-306-0005)
Storage Solution 100 mL (GP-435-0100)
Eppendorf LoBind® 0.5 mL tubes package of 50 (C275-130)

Specifications

Specifications Description
Sample Volume Required 2 µL
Total Electrophoresis Run Time 50 minutes (22-40 Array)
DNA Sizing Range 100 bp – 6,000 bp
Separation Resolution 100 bp -1,200 bp = 5%
1,200 bp – 6,000 bp = 15%
DNA Sizing Accuracy ± 5%
DNA Sizing Precision 2% CV
DNA Fragment Detection Range (S/N > 5) 0.05 pg/µL – 5 pg/µL
DNA Smear Detection Range (S/N > 5) 5 pg/µL – 250 pg/µL
DNA Fragment Quantification Range (S/N > 10) 0.1 pg/µL – 5 pg/µL
DNA Smear Quantification Range (S/N > 10) 25 pg/µL – 250 pg/µL
DNA Quantification Accuracy ± 25%
DNA Quantification Precision 15% CV
Maximum DNA Concentration 5 pg/µL per fragment; 250 pg/µL total

Examples

The precision sizing and high-sensitivity DNA detection provided by the Ultra-Sensitivity NGS Analysis Kit (FP-1101-0275) depends in part upon the employment of a carefully designed DNA ladder. The Ultra-Sensitivity NGS Ladder (FP-7101-U060) is comprised of DNA fragments at precisely measured concentrations to enable precision sizing and sensitive quantification.

Figure 1
Figure 1. Separation of the Ultra-Sensitivity NGS Ladder (FP-7101-U060) performed on a FEMTO Pulse™ Automated Pulsed-Field CE Instrument equipped with an Ultra-Short Capillary Array (22-40). Separations performed with the Ultra-Sensitivity NGS Analysis Kit (FP-1101-0275).

Quality control analysis in NGS library workflows is a crucial step to ensure superior sequencing results. The Ultra-Sensitivity NGS Analysis Kit (FP-1101-0275) facilitates the detection of NGS smears down to
5 pg/µL. NGS smears are quantified through 250 pg/µL (Figure 2). Such sensitive nucleic acid detection enables researchers to conserve precious sample across crucial QC checkpoints for further downstream analysis. Beyond conserving precious samples in typical NGS workflows, the Ultra-Sensitivity NGS Analysis Kit has various applications including: single cell genomics, PCR-free NGS library construction, metagenomics, and metatranscriptomics, among others.

Figure 2
Figure 2. The upper panel depicts the electropherogram of a commercially prepared NGS library diluted to 5 pg/µL. The bottom panel depicts the electropherogram of a commercially prepared NGS library diluted to 250 pg/µL. Separations were performed on a FEMTO Pulse Automated Pulsed-Field CE Instrument equipped with an Ultra-Short Capillary Array (22-40) using the Ultra-Sensitivity NGS Analysis Kit (FP-1101-0275).

The DNA fragment detection sensitivity is similarly impressive. The DNA fragment detection range is expansive, covering from 50 fg/µL to 5 pg/µL (Figure 3).

Figure 3
Figure 3. The upper panel depicts the electropherogram of a 300 bp DNA fragment diluted to 60 fg/µL. The bottom panel depicts the electropherogram of a 300 bp DNA fragment diluted to 5 pg/µL. Separations were performed on a FEMTO Pulse Automated Pulsed-Field CE Instrument equipped with an Ultra-Short Capillary Array (22-40) using the Ultra-Sensitivity NGS Analysis Kit (FP-1101-0275).

Post-separation analysis is expedited with PROSize™ Data Analysis Software, which reports on calculated DNA fragment and DNA smear size and concentration. Furthermore, the Smear Analysis function provides researchers with a tool to more accurately size and quantify NGS smears for downstream analysis.

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