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

High Sensitivity NGS Fragment Analysis Kit

High Sensitivity NGS Fragment Analysis Kit
500 Sample High Sensitivity NGS Fragment Analysis Kit

The High Sensitivity NGS Fragment Analysis Kit provides an automated solution to the quality assessment of low concentration NGS libraries prior to sequencing. The correct determination of molarity for optimum flow-cell loading is a critical step in NGS, and DNF-474 efficiently and accurately determines the average size of a library and its concentration. The correct calculation of molarity helps ensure quality NGS results. Use DNF-474 to increase the efficiency of your genomic research.

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Components

Components 500 Sample Kit 1,000 Sample Kit
NGS Fragment Separation Gel 240 mL (DNF-240-0240) 500 mL (DNF-240-0500)
Intercalating Dye 30 µL (DNF-600-U030) 30 µL x 2 (DNF-600-U030)
5X 930 dsDNA Inlet Buffer 125 mL (DNF-355-0125) 300 mL (DNF-355-0300)
5X Capillary Conditioning Solution 50 mL (DNF-475-0050) 100 mL (DNF-475-0100)
HS NGS Fragment Diluent Marker Solution 2.4 mL x 5 vials (DNF-373-0003) 2.4 mL x 10 vials (DNF-373-0003)
HS NGS Fragment DNA Ladder 100 µL (DNF-396-U100) 100 µL x 2 (DNF-396-U100)
0.25X TE Rinse Buffer 125 mL (DNF-497-0125) 125 mL (DNF-497-0125)
BF-25 Blank Solution 8 mL (DNF-300-0008) 8 mL (DNF-300-0008)

Specifications

Specifications Description
Sample Volume Required 2 µL
Diluent Marker Volume Required 22 µL
DNA Sizing Range 100 bp – 6,000 bp
Separation Resolution Short Array: 25 bp - 100 bp ≤ 10%; 100 bp - 2000 bp ≤ 5%; 2000 bp - 5000 bp ≤ 10%
DNA Sizing Accuracy ± 5% or better
DNA Sizing Precision 2% CV
DNA Fragment Concentration Range 5 pg/µL – 500 pg/µL input DNA
DNA Smear Concentration Range 50 pg/µL – 5000 pg/µL input DNA
DNA Quantification Accuracy ± 25 %
DNA Quantification Precision 15 % CV
Maximum DNA Concentration 500 pg/µL per fragment; 5000 pg/µL total
Total Electrophoresis Run Time 25 min (Ultra-Short Array, 22-47); 50 min (Short Array, 33-55); 80 min (Long Array, 55-80)

Examples

In the following section we have provided several examples of typical and atypical separations performed with the HS NGS Fragment Analysis Kit on a Fragment Analyzer fitted with a standard short array.

Figure 1 depicts a typical separation of the HS NGS DNA Ladder (DNF-396). The peaks are clearly defined and well separated, providing a precise and accurate standard curve for the sizing and quantitation of samples.

Figure 1
Figure 1. A typical separation of the HS NGS DNA Ladder (DNF-396) on a Fragment Analyzer equipped with a Short Capillary Array (33-55) using the High Sensitivity NGS Fragment Analysis Kit (DNF-474).

Assessment of the size and quality of a library smear is important to obtaining quality sequencing results. Figure 2 shows a high quality library constructed from commercially prepared human genomic DNA.

Figure 2
Figure 2. A high quality NGS library. Samples were run using the HS NGS Fragment Analysis Kit on a Fragment Analyzer equipped with a Short Capillary Array (33-55).

Importantly, the HS NGS Fragment Kit is able to detect primer dimers and adapter dimers, significant indicators of library quality. High levels of dimers, adapter or primer, interferes with sequencing, reducing the quality of the results. Figure 3 depicts a separation of an NGS library that has primer dimers and adapter dimers. Primer dimers typically appear around 70 bp, while adapter dimers appear around 140 bp.

Figure 3
Figure 3. An NGS library with primer and adapter dimers. Samples were run using the HS NGS Fragment Analysis Kit on a Fragment Analyzer equipped with an Ultra-Short Capillary Array (22-47).
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