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

Mutation Discovery 910 Gel Kit
Mutation Discovery 920 Gel Kit

Mutation Discovery 910 Gel Kit (DNF-910T) and Mutation Discovery 920 Gel Kit (DNF-920T)
Mutation Discovery 910 Gel Kit
Mutation Discovery 920 Gel Kit

The Mutation Discovery Kits, when employed with the dsDNA Cleavage Kit (DNF-480), provide researchers with an efficient TILLiNG discovery protocol. A heteroduplex cleavage strategy is employed to efficiently identify point and small mutations such as SNPs. Each kit is provided with a unique, specialized gel formulation. The Mutation Discovery 910 Gel Kit (DNF-910T) uses a gel formulation best suited to DNA fragments below 1,000 bp, while the Mutation Discovery 920 Gel Kit (DNF-920T) was developed for DNA fragments between 1,000 bp and 3,000 bp. Note: These kits have been optimized for use with a 96 capillary array.

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Components

Components 1,000 Sample Kit (DNF-910T) 1,000 Sample Kit (DNF-920T)
dsDNA Gel 35 bp – 1,500 bp 500 mL (DNF-810-0500) 500 mL (DNF-920-0500)
Intercalating Dye 30 µL x 2 (DNF-600-U030) 30 µL x 2 (DNF-600-U030)
5X 930S dsDNA Inlet Buffer 300 mL (DNF-355-0300) 300 mL (DNF-455-0300)
5X Capillary Conditioning Solution 100 mL (DNF-475-0100) 100 mL (DNF-475-0100)
Mutation Detection DNA Ladder 1.0 mL (FS-SLR480-0001) 1.0 mL (FS-SLR480-0001)
35 bp and 5,000 bp Markers 3.2 mL (FS-SMK480-0003) 3.2 mL (FS-SMK480-0003)
Mineral Oil Dropper Bottle 15 mL (FS-SMO15) 15 mL (FS-SMO15)

Specifications

Specifications DNF-910T DNF-920T
Sample Volume Required 2 µL
Diluent Marker Volume Required 22 µL
DNA Sizing Precision 2% CV
DNA Fragment Concentration Range 5 ng/µL – 100 ng/µL input DNA
Total Electrophoresis Run Time 50 min (Short Array); 80 min (Long Array) 45 min (Short Array); 75 min (Long Array)

Examples

Identification of TILLiNG induced mutations necessitates a well separated, quality DNA ladder for sizing samples. The Mutation Detection DNA Ladder (FS-SLR480-0001) provides accurate sizing from 35 bp – 5,000 bp via well separated ladder fragments and markers. The ladder enables the necessary sizing distinction required to efficiently separate enzyme mismatch assay products.

Figure 1
Figure 1. Separation of the Mutation Detection DNA Ladder (FS-SLR480-0001) performed on a Fragment Analyzer equipped with a Short Capillary Array (33-55) using the Mutation Discovery 910 Gel Kit (DNF-910T).

The Mutation Discovery Gel Kits (DNF-910T, DNF-920T) provides an efficient automated solution to identifying mutant amplicons after an enzyme mismatch (heteroduplex) assay has been performed. Traditionally, reaction products would be separated using agarose gel electrophoresis. While a cost-effective approach agarose gel electrophoresis is labor intensive and lacks the resolving power of the Fragment Analyzer. Separation of the products from the enzymatic mismatch assay using the Mutation Discovery Gel Kits (DNF-910T, DNF-920T) allows for the efficient and concurrent electrophoresis of up to 96 samples, as shown in Figure 2.

Figure 2
Figure 2. Digital gel image of 96 TIILiNG samples separated with the Mutation Discovery 920 Gel Kit (DNF-920T) on a Fragment Analyzer equipped with a Short Capillary Array (33-55).

Post-electrophoretic analysis in PROSize® simplifies the identification of wild-type and mutant amplicons, as shown in Figure 3.

Figure 3
Figure 3. Separation of a positive mutation sample using the Mutation Discovery 910 Kit (DNF-910T). Electrophoresis was performed on a Fragment Analyzer equipped with a Short Capillary Array (33-55). The parent amplicon peak is highlighted red and the two daughter peaks are highlighted orange.

If an amplicon does not contain a mutation, cleavage will not occur and only the parent amplicon will be present, as depicted in Figure 4. However, if an amplicon contains a mutation, PROSize will automatically highlight the parent amplicon red and the cleavage products orange, accelerating mutation discovery.

Figure 4
Figure 4. Separation of a sample negative for a mutation using the Mutation Discovery 920 Gel Kit (DNF-920T). Electrophoresis was performed on a Fragment Analyzer equipped with a Short Capillary Array (33-55). Only the parent amplicon is present.
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