3`RNA End Labeling 647
Kit to Indroduce a Single Modification at the 3'END of your mRNA
- Catalog No.
- 1 Kit
- 760 €
This 3’ RNA End Labeling Kit has been developed for in vitro labeling of mRNA molecules. The kit introduces a single azide at the 3’-end of the mRNA. This modification strategy ensures good translation efficieny of the unmodified open reading frame (ORF). The azide at the poly(A) tail can be reacted with alkyne-derivates of fluorescent dyes, haptenes and other labels in a highly selective fashion under benign click reaction conditions. Finally, fluorescently labeled mRNA can be used to detect localization of the mRNA inside of cells after transfection.
S.Croce, S. Serdjukow, T. Carell, T. Frischmuth, ChemBioChem, 2020, 21, 1-7
Can the 3’ END labelling kit be used with other RNAs?
Indeed, it can be used for other kinds of RNA. Make sure that your RNA does not have secondary structures that can interfere with enzyme activity. If you are not sure about secondary structures, it can help to heat your samples shortly (5 minutes at 65 °C and then on ice for other 5 minutes) before applying the reagents. We recommend to try such a step first with a smaller test sample, as too harsh handling can also affect your RNA. Importantly, make sure that your RNA is longer than 100 nucleotides, since the purification method provided with the kit will not be able to retain shorter fragments.
Does my RNA need to end with an A nucleotide for efficient 3’ END labelling?
No this is not necessary, the poly(A) polymerase inside the kit is able to add the modification whatever the last natural base is.
Which kit should be used for RNA purification?
RNA purification can be achieved using spin columns purification methods such as RNeasy MinElute Cleanup Kit from Qiagen.
What is the best way to handle RNA to avoid degradation?
Due to its chemical structure RNA is less stable than DNA. At high temperatures, under basic conditions (pH>9) and in the presence of high Mg2+ concentrations, chemical reactions that favour hydrolysis of the RNA molecule take place. Additionally, RNases are common contaminations in standard laboratory conditions compared to DNases. We recommend to clean the benches and equipment with RNase inhibitors or specific detergent mixtures and to wear fresh gloves when handling the tubes (gloves that have been worn or that e.g. touched door/refrigerator handles are considered contaminated). Moreover, RNase-free H2O and tubes should be used for buffer preparation and to dissolve the samples.
What is the best way to store RNA to avoid degradation?
For storage we recommend to store RNA at –80 °C, which should conserve RNA for one year without significant degradation. Alternatively.
In our experience, short synthetic RNAs (up to 50mer) are more stable than long single-stranded RNAs from biological samples and therefore can be stored at –20 °C in a dry format.
- Can the 3’ END labelling kit be used with other RNAs?
12 months unopened after receipt
diverse, see labels on the kit
kit system made of different components
please see user manual of the kit
- Shelf Life