BaseClickTM-Kit Eterneon 350/455 Protocol - Jena Bioscience
BaseClickTM-Kit Eterneon 350/455 Protocol - Jena Bioscience
BaseClickTM-Kit Eterneon 350/455 Protocol - Jena Bioscience
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BaseClick TM -<strong>Kit</strong> (Oligonucleotide and DNA Labeling <strong>Kit</strong>)<br />
baseclick GmbH<br />
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• The reaction time depends on: a) concentration of azide and oligo in the solution;<br />
b) reaction temperature; c) stirring and/or mixing of the solution.<br />
• Excess of reagents are normally removed by ethanol precipitation of the labeled<br />
oligonucleotide.<br />
B. Materials provided with the <strong>Kit</strong> and storage conditions<br />
Vialcolour<br />
Amount Component Storage<br />
red 1 mg <strong>Eterneon</strong> TM <strong>350</strong>/<strong>455</strong> Azide 2 dark, -20 °C<br />
yellow 5,4 mg TBTA-Ligand 1, 2 -20 °C<br />
green 6 x 1 mg CuBr 1 dry, inert gas<br />
blue 2 x1 mL DMSO/t-BuOH 1 RT<br />
1 baseclick grade<br />
2 a prepared solution can be stored at - 20 °C in the dark for several months<br />
(Note: The azide functionality is very stable and does not hydrolyze in the presence of water.)<br />
C. Required Material and Equipment<br />
Alkyne-modified oligonucleotide (can be ordered @ baseclick GmbH, Integrated DNA<br />
Technologies, Metabion, IBA or EllaBiotech)<br />
Microcentrifuge<br />
Vortexer<br />
Thermomixer (optional)<br />
Microcentrifuge tubes<br />
Ethanol (5 % Diethylether)<br />
0,3 M Sodium-acetate solution (0,3 M NaOAc)<br />
D. Click-<strong>Protocol</strong> for Oligonucleotide Labeling<br />
1. Before You Start<br />
a) Calculate the total amount of terminal alkyne modifications n in nmol present in your<br />
oligonucleotide:<br />
n = m x y where:<br />
n = terminal alkyne modifications in nmol<br />
m = amount of oligonucleotides in nmol<br />
y = number of terminal alkyne functions in your oligonucleotide<br />
e.g. you have 20 nmol of a triple alkyne modified oligonucleotide:<br />
amount of terminal alkyne modifications n = 20 nmol x 3 = 60 nmol.<br />
2