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

Standard Sensitivity Large Fragment Analysis Kit

Standard Sensitivity Large Fragment Analysis Kit
500 Sample Standard Sensitivity Large Fragment Analysis Kit

The Standard Sensitivity Large Fragment Analysis Kit is used for the automated sizing and quantification of large DNA fragments and smears. Common applications include as a quality assurance step in long read sequencing, checking the quality of a DNA extraction, and evaluating the quality of degraded DNA samples. Ideal for high concentration samples, this kit is able to size samples through 20 Kb.

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Components

Components 500 Sample Kit 1,000 Sample Kit
Large Fragment Separationg Gel 240 mL (DNF-220-0240) 500 mL (DNF-220-0240)
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)
SS Large Fragment Diluent Marker Solution 2.4 mL x 5 vials (DNF-380-0003) 2.4 mL x 10 vials (DNF-380-0003)
SS Large Fragment DNA Ladder 100 µL (DNF-378-U100) 100 µL x 2 (DNF-378-U100)
0.6X TE Rinse Buffer 125 mL (DNF-496-0125) 125 mL (DNF-496-0125)
BF-2000 Blank Solution 8 mL (DNF-302-0008) 8 mL (DNF-302-0008)

Specifications

Specifications Description
Sample Volume Required 2 µL
Diluent Marker Volume Required 22 µL
DNA Sizing Range 50 bp – 20,000 bp
DNA Sizing Accuracy ± 15% or better
DNA Fragment Concentration Range 0.1 ng/µL - 10 ng/µL input DNA
DNA Smear Concentration Range 5 ng/µL - 100 ng/µL input DNA
DNA Quantification Accuracy ± 25%
DNA Quantification Precision 20% CV
Maximum DNA Concentration 10 ng/µL per fragment; 100 ng/µL total
Total Electrophoresis Run Time 50 minutes (Short Array, 33-55); 80 minutes (Long Array, 55-80)

Examples

A typical separation of the Standard Sensitivity Large Fragment Ladder is depicted in Figure 1. A high quality ladder separation is crucial to the accurate quantification and sizing of large DNA fragments. The well separated ladder fragments allow for the accurate sizing of large DNA fragments through 20,000 bp and accurately quantifies fragments as low as 0.1 ng/µL.

Figure 1
Figure 1. A typical separation of the Standard Sensitivity Large Fragment DNA Ladder (DNF-378) on a Fragment Analyzer equipped with a Short Capillary Array (33-55) using the Standard Sensitivity Large Fragment Analysis Kit (DNF-492).

Through the use of high quality reagents and carefully selected electrophoresis parameters, the Standard Sensitivity Large Fragment Analysis Kit (DNF-492) is able to accurately measure large DNA fragment through 20 Kb. Figure 2 depicts the separation of a synthetic 15 Kb DNA fragment using this kit. In this example, PROSize® measured this fragment to within 3% of its known size.

Figure 2
Figure 2. A synthetic 15 Kb DNA fragment separated on a Fragment Analyzer equipped with a Short Capillary Array (33-55) using the Standard Sensitivity Large Fragment Analysis Kit (DNF-492).

Efficient and accurate sizing of large DNA fragments is a significant aspect of many molecular biology applications including next generation long read sequencing, assessment of purification success, and generation of large synthetic gene constructs, among others. Notably, assessment of large DNA fragments is crucial for Human Leukocyte Antigen (HLA) typing, a process essential for matching donor organ and tissue to recipients. Click here to learn more about how the Anthony Nolan Research Institute uses the large fragment kits for the Fragment Analyzer in HLA typing workflows.

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