HighFidelity RED PCR Labeling Testkit

HighFidelity RED PCR Labeling Testkit is designed to produce randomly Cy5-, AF647-and ATTO643-modified DNA probes by PCR to find the optimal label for the red emission wavelength range. Such probes are ideally suited for Fluorescence in situ hybridization (FISH) and Northern Blot experiments. PCR-based labeling is superior to random-primed labeling with Klenow fragment if template amounts are limited or amplification of a specific DNA fragments is required. Amplification of probes up to 4 kbp is feasible. All labeled-dUTP are efficiently incorporated into DNA as sub-stitute for its natural counterpart dTTP using an optimized reaction buffer and a High Fidelity Polymerase blend consisting of Taq polymerase and a proofreading enzyme. 50% labeled-dUTP substi-tution typically results in an optimal balance between reaction and labeling efficiency. Individual optimization of labeled-dUTP/dTTP ra-tio however, can easily be achieved with the single nucleotide format.

Product Specifications

Product Name Alternative

Preparation of Cy5-, AZDye647- and ATTO643-labeled DNA probes by PCR

Field of Research

Molecular Biology

Storage Conditions

Store at -20 °C

Notes

For research use only.

Applications Notes

Spectroscopic Propertie: Cy5: λexc 649 nm, λem 670 nm, ε 250.0 L mmol-1 cm-1 (Tris-HCl pH 7.5), AF647: λexc 648 nm, λem 671 nm, ε 270.0 L mmol-1 cm-1 (Tris-HCl pH 7.5), ATTO643: λexc 643 nm, λem 665 nm, ε 150.0 L mmol-1 cm-1 (Tris-HCl pH 7.5) .