Label IT^® Nucleic Acid Labeling Kit, CX-Rhodamine – labels 25 µg nucleic acid

Supporting Data

Label IT® Nucleic Acid Labeling Kits. The Label IT® chemical labeling reagents are composed of three regions: the label (fluorophore or hapten) (green), the linker (yellow) which facilitates electrostatic interactions with nucleic acids and the reactive alkylating group (blue) that covalently attaches the Label IT® reagent to any reactive heteroatom within the nucleic acids. Attachment of the Label IT® Reagents to nucleic acids does not alter the structure of the nucleic acid or affect downstream hybridization performance, and as such, nucleic acids labeled using the Label IT® Reagents can be employed in multiple applications as defined by the researcher.

Non-destructive Direct Labeling of Nucleic Acid. Mouse total RNA was labeled with Label IT® Cy®3 (B) electrophoresed and analyzed alongside unlabeled total RNA (A) on an Agilent RNA 6000 Nano chip. RNA Integrity Number (RIN) is approximately 9.0 for both labeled and unlabeled samples.

Labeling Density can be Easily Controlled with the Label IT® Reagents. Plasmid DNA was labeled with increasing ratios (w:w) of Label IT® CX-Rhodamine to DNA. The extent of labeling was estimated, and normalized to the amount of recovered DNA (# labels/100 base pairs), using two different criteria: fluorescence intensity of the attached rhodamine (excitation at 585 nm, emission at 610 nm), and visible absorbance at 585 nm. The significance of quenching, at the higher labeling ratios, becomes apparent in the fluorimetric estimations of labeling density.