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

Small RNA Analysis Kit

Small RNA Analysis Kit
275 Sample Small RNA Analysis Kit

The Small RNA Analysis Kit provides automated analysis of small RNAs. The kit excels in the evaluation of small RNAs for downstream applications, notably including small RNA NGS libraries. Able to distinguish between precursor small RNA and functional small RNA, it can also quantify the amount of microRNA present in any given sample. To help streamline laboratory workflows, post-separation analysis using the Small RNA Analysis Region function in PROSize® 2.0 easily quantifies the microRNA and small RNA present in a sample. Note: This kit is optimized for use on a 12 capillary array.

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Components

275 Sample Kit
Small RNA Separation Gel 250 mL (DNF-262-0250)
Intercalating Dye 30 µL (DNF-600-U030)
5X 930 dsDNA Inlet Buffer 125 mL (DNF-355-0125)
5X Capillary Conditioning Solution 50 mL (DNF-475-0050)
Small RNA Diluent Marker 4 mL x 2 (DNF-368-0004)
Small RNA Ladder 60 µL (DNF-361-U060)
0.25X TE Rinse Buffer 125 mL (DNF-497-0125)
BF-25 Blank Solution 8 mL (DNF-300-0008)
Eppendorf LoBind® 0.5 mL tubes package of 50

Specifications

Specifications Description
Sample Volume Required 2 µL
Diluent Marker Volume Required 18 µL
RNA Sizing Range 5 - 200 nt
Qualitative Range 25 pg/µL - 2500 pg/µL (microRNA region)
Quantitative Range 50 pg/µL - 2000 pg/µL (microRNA region)
Quantification Precision 25% CV (Small RNA Ladder)
Total Electrophoresis Run Time 17 minutes (Ultra-Short Array, 22-47); 22 minutes (Short Array,33-55)

Examples

With the increasing interest in small RNAs for downstream applications such as NGS, the accurate and efficient analysis of small RNAs from a variety of sources is crucial. Small RNA sample analysis is dependent upon the employment of a high quality small RNA ladder as seen in Figure 1. The Small RNA Analysis Kit (DNF-470) is able to measure the size and concentration of small RNAs, including functional microRNAs, due to the Small RNA Ladder (DNF-361). The Small RNA Ladder, comprised of small RNA fragments, enhances the accuracy and precision of fragment size and concentration measurements beyond other analysis methods.

Figure 1
Figure 1. A typical separation of the Small RNA Ladder (DNF-361) performed on a Fragment Analyzer equipped with a Short Capillary Array (33-55) using the Small RNA Analysis Kit (DNF-470).

The Small RNA Analysis Kit (DNF-470) provides a more detailed analysis of the small RNA region than possible with the HS Total RNA Analysis Kit (DNF-472) as shown in Figure 2. The same total RNA sample from Corn Leaf was run on the Fragment Analyzer using each respective analysis kit. The HS Total RNA Analysis Kit (DNF-472) provides an excellent overview of the sample, indicating that the extracted total RNA is high quality with an RQN of 10.0. The Small RNA Analysis Kit (DNF-470) takes the analysis a step further with not only a more detailed separation of the small RNA region, but quantifying the amount of small RNA and microRNA present in the total RNA sample.

Figure 2a
Figure 2b
Figure 2. Total RNA extracted from corn leaf separated using the HS Total RNA Analysis Kit (15 nt) (DNF-472) (TOP) and the Small RNA Analysis Kit (DNF-470) (BOTTOM). Capillary electrophoresis was performed on a Fragment Analyzer equipped with a Short Capillary Array (33-55). Both kits show the small RNA region, but the Small RNA Analysis Kit (DNF-470) depicts the region in more detail, distinguishing between the microRNA region (10 nt – 40 nt) and the small RNA region (10 nt – 200 nt).
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