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

Ultra-Sensitivity RNA Analysis Kit

Analysis of RNA is a crucial aspect of many workflows in genomics and molecular biology. The Ultra-Sensitivity RNA Analysis Kit (FP-1201-0275) facilitates efficient and accurate quality analysis of RNA with unprecedented sensitivity. Qualify and quantify low concentration transcripts and conserve precious sample with the Ultra-Sensitivity RNA Analysis Kit and the FEMTO Pulse™ Automated Pulsed-Field CE Instrument.

Components

275 Sample Kit
RNA Separation 5201 Gel 250 mL (FP-5201-0250)
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 RNA Diluent Marker 3 mL x 2 vials (FP-8201-0003)
Ultra-Sensitivity RNA Ladder 60 µL (FP-7201-U060)
RNA Diluent Buffer 3 mL (FP-6501-0003)
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 45 minutes (22-40 Array)
RNA Fragment Sizing Accuracy ± 20%
RNA Fragment Sizing Precision 20% CV
RNA Detection Range 2.5 pg/µL – 250 pg/µL
RNA Quantification Range 15 pg/µL – 250 pg/µL
RNA Quantification Accuracy ± 30%
RNA Quantification Precision 20% CV

Examples

Quantification and qualification of RNA with the FEMTO Pulse necessitates in part the use of a high quality RNA ladder. A well designed RNA ladder provides robust standard curves for both fragment sizing and sample quantification. The Ultra-Sensitivity RNA Ladder (FP-7201-U060) is comprised of carefully selected RNA fragments aliquoted in precise concentrations (Figure 1).

Figure 1
Figure 1. Separation of the Ultra-Sensitivity RNA Ladder (FP-7201-U060) performed on the FEMTO Pulse Automated Pulsed-Field CE Instrument equipped with an Ultra-Short Capillary Array (22-40). PFCE performed with the Ultra-Sensitivity RNA Analysis Kit (FP-1201-0275).

Quantification and qualification of total RNA samples are crucial steps for various downstream applications, notably as a raw materials QC for various NGS workflows. The Kit is capable of evaluating RNA from all biological sources: plant, animal, microbial, and fungal. The unprecedented sensitivity provided by the Ultra-Sensitivity RNA Analysis Kit (FP-1201-0275) enables researchers to both conserve precious sample and evaluate RNA at low sample concentration (Figure 2).

Figure 2
Figure 2. The upper panel shows the electropherogram of a mouse total RNA separation at the lower end of the kit quantification range. The lower panel shows the electropherogram of a rice leaf total RNA separation at the bottom of the quantification range. Capillary electrophoresis was performed on a FEMTO Pulse Automated Pulsed-Field CE Instrument equipped with an Ultra-Short Capillary Array (22-40) using the Ultra-Sensitivity RNA Analysis Kit (FP-1201-0275).

Post-capillary electrophoresis analysis is simplified and expedited in the PROSize™ Data Analysis Software through the use of the RNA Quality Number (RQN), reported in the RNA Property Summary table (Figure 1). A quantitative quality metric developed by scientists at Advanced Analytical, the RQN assigns RNA samples a numeric score between 0 and 10. As with most quality metrics, the RQN exists on a continuum; samples with an RQN of 7 or higher are generally considered ideal for downstream applications. An RQN below 7 indicates issues with the RNA sample, including: degradation, contamination, and poor extraction and purification results.

The importance of good RNA quality for RNA-Seq and other downstream applications is quickly illustrated by performing a degradation series and analyzing the results on the FEMTO Pulse (Figure 3). As samples are degraded, a corresponding decrease in the RQN is observed. On the electropherogram this is shown as a decrease in the intensity of the ribosomal peaks and the appearance of a degradation smear. The end result is a loss of intact RNA, decreasing the efficiency of NGS applications.

Figure 3
Figure 3. A degradation series of total RNA extracted from Arabadopsis thaliana. Total RNA samples were degraded at 90° C with an aliquot collected at 2, 4, and 6 minutes. Capillary electrophoresis was performed on a FEMTO Pulse Automated Pulsed-Field CE Instrument equipped with an Ultra-Short Array (22-40) using the Ultra-Sensitive RNA Analysis Kit (FP-1201-0275).
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