RNase Contamination Assay Kit
- This product was discontinued on 01/01/2016
Evaluation of RNase contamination is necessary for reagents to be used in experiments with RNA. The RNase Contamination Assay Kit detects general RNase activities including non-enzyme based RNA degradation due to heavy metal contamination in samples and high pH. The assay probe is a fluorescein labeled RNA transcript (300-mer). After incubation with a reagent sample the integrity of the RNA probe is analyzed on denaturing PAGE followed by SYBR Gold staining or preferably by scanning with a FAM/Fluorescein capable imaging system like the Typhoon scanner. Most samples (e.g. enzymes, column fractions from enzyme purifications, unlabeled RNA/DNA samples, buffers, or diluents) can be tested directly. The assay is semi-quantitative and is more sensitive if detergent is present.
In addition to RNase assay, the labeled RNA probe can also be used in studies involving Poly(A) and Poly(U) tailing, RNA ligation, RNA capping/decapping assays and sequence and structure specific RNase assays.
Materials not included:
Nuclease-free water, incubator, PAGE/Urea gel, TBE running buffer, power supply, SYBR Gold, Typhoon scanner and UV light box.
Fluorescein Labeled RNA Probe:
The probe is an in vitro RNA transcript (300-mer), internally labeled with Fluorescein-12-UTP, supplied at 40 ng/µl.
1 GGGAAGAGCA UUGUCGAGGG UGAAGUACGG UUCUUGCAGG UUGAACAUCA GCGCGCUCUU
61 ACCUUUCGCU UUCAGUUCUU UAUCCAGCGC CGGGAUCUCU UCCCAGGUUU UUGGCGGGUU
121 CGGCAGCAGA UCUUUGUUAU AAAUCAGCGA UAACGCUUCA ACAGCGAUCG GGUAAGCAAU
181 CAGCUUGCCG UUGUAACGUA CGGCAUCCCA GGUAAACGGA UACAGCUUGU CCUGGAACGC
241 UUUGUCCGGG GUGAUUUCAG CCAACAGGCC AGAUUGAGCG UACUCGACGA AGACUCCCUA
1X NEBuffer 4:
50 mM Potassium acetate
20 mM Tris-acetate
10 mM Magnesium acetate
1 mM Dithiothreitol
(pH 7.9 at 25°C)
2X RNA Loading Dye:
0.02% Bromophenol blue
0.01% Xylene cyanol
1 mM EDTA
The following reagents are supplied with this product:
Store at (°C) Concentration RNA Loading Dye, (2X) -20 2 X NEBuffer™ 4 -20 10 X Fluorescein-RNA Probe 40 μg/ml Proteinase K, Molecular Biology Grade -20 800 units/ml
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