PCR DIG Probe Synthesis Kit

The PCR DIG Probe Synthesis Kit contains an alkali-labile digoxigenin (DIG)-11deoxyuridinetriphosphate (dUTP) formulation. This kit is for convenient and efficient generation of DIG-labeled DNA probes that are highly sensitive in polymerase chain reaction (PCR). The probes are labeled with DIG-11dUTP (alkali-labile), by the method of PCR. The ratio of DIG-dUTP:dTTP is 1:2.
The PCR DIG Probe Synthesis Kit contains the Expand High Fidelity DNA polymerase mix. This robust enzyme mix with proofreading activity will polymerize probes 40 bp to 5 kb long using 10 pg plasmid DNA and 10 ng genomic DNA as template. The DIG-labeled probes are stable for over one year.

Digoxigenin (DIG)-labeled DNA probes produced from the PCR DIG Probe Synthesis Kit is suitable for low-(single)-copy gene detection of rare mRNA in Southern and northern blots. DIG-labeled DNA probes have also been used for labeling of DNA during in situ hybridization.
The concentration of the supplied dUTP-nucleotide mix can be adjusted according to probe length. Labeling effectiveness can quickly be determined on an agarose gel.
Stripping and reprobing of membranes is possible multiple times following the protocol in the package insert.
One PCR labeling reaction (50 μl) will typically yield enough probe for 20 ml hybridization solution. The kit can be used for approximately 25 reactions (50 μl).

1 kit containing 6 components.

Each lot of the PCR DIG Probe Synthesis Kit is function tested in PCR. Amplification products are assayed in genomic Southern blots.
Under PCR conditions as described in this package insert, the control reaction generates an amplification product of 442bp. Due to multiple incorporations of DIG-dUTP during the PCR process, the molecular weight of the PCR products is significantly increased compared to the unlabeled PCR product. A specific fragment pattern is detected after hybridization of the PCR product to 10μg human genomic DNA followed by chemiluminescent detection.