The Ambion® BrightStar® Psoralen-Biotin Nonisotopic Labeling Kit is for the biotin labeling of nucleic acids. Probes generated by this method have the maximum amount of biotin incorporated that will not interfere with blot hybridization, yielding maximum sensitivity. The kit includes sufficient reagents for 20 reactions.
• Simple, scalable, single-reagent protocol
• Very high sensitivity (<100 fg), comparable to radiolabeled probes
• Versatile—one kit labels RNA, DNA, PCR products, cDNA inserts, and oligonucleotides
• Chemical incorporation allows labeling of already prepared nucleic acids
One-Step Chemical Labeling
BrightStar® Psoralen-Biotin consists of a nucleic acid intercalating moiety, psoralen, covalently attached to biotin. The psoralen-biotin intercalates within the nucleic acid (single- or double-stranded) and is then covalently bound by irradiation with long-wave UV light.
Synthesize Probes Just Once
The entire labeling reaction takes only 45 min. 0.5 µg of template will generate sufficient probe for 50 mL of traditional northern or Southern hybridization solution or for 500 mL of ULTRAhyb® Ultrasensitive Hybridization Buffer. The reaction is scalable, which allows the production of large mass amounts of nonisotopically labeled probes in a single reaction. Since these probes are not subject to radiolysis as are radiolabeled probes, they may be stored for at least a year.
One Kit Labels Any Nucleic Acid
Since BrightStar® Psoralen-Biotin chemically labels nucleic acids that are already prepared by conventional methods, the reaction is not dependent on the amount and purity of the template, as are enzymatic labeling methods. This means that labeling reactions using BrightStar® Psoralen-Biotin are easier to perform and are more consistent and robust than other nonisotopic labeling methods. It can be used to label single-stranded antisense RNA for northerns, Southerns, or in situ hybridizations. The same kit can also label PCR products, plasmids, gel purified cDNA inserts, and oligonucleotides for northern and Southern blots and in situ hybridizations. Sensitivities are comparable to those obtained with 32P radiolabeled probes and enzymatically synthesized biotin-labeled probes when used in conjunction with the BrightStar® BioDetect™ Nonisotopic Detection Kit and BrightStar®-Plus membrane (see Accessory Products below). It is not recommended to label probes with this kit for use with ribonuclease protection assays or S1 assays. Instead, an enzymatic nonisotopic labeling method should be used.
For the highest sensitivity possible, use the BrightStar® Psoralen-Biotin Kit in conjunction with BrightStar® -Plus Positively Charged Nylon Membranes (SKU# AM10100, AM10102, or AM10104) and the BrightStar® BioDetect™ Nonisotopic Detection Kit (SKU# AM1930). For the visualization and gel purification of psoralen-biotin labeled probes use the Fluor-coated TLC Plates (SKU# AM10110) and the Portable UV Light (SKU# AM12750).