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11636090910

Roche

PCR DIG Probe Synthesis Kit

sufficient for 25 reaction (50 μL final reaction volume)

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Synonym(s):
DIG system, probe

usage

sufficient for 25 reaction (50 μL final reaction volume)

Quality Level

manufacturer/tradename

Roche

technique(s)

PCR: suitable

storage temp.

−20°C

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This Item
115855509101158561491011585606910
PCR DIG Labeling Mix solution, suitable for PCR

Roche

11585550910

PCR DIG Labeling Mix

DIG-High Prime sufficient for 40 labeling reactions, pkg of 160 μL, solution

Roche

11585606910

DIG-High Prime

usage

sufficient for 25 reaction (50 μL final reaction volume)

usage

sufficient for 2 x 25 assays (100 ul final reaction volume)

usage

sufficient for 12 labeling reactions (10 ng to 3 μg per assay), sufficient for 24 blots (blots of 100 cm2)

usage

sufficient for 40 labeling reactions

manufacturer/tradename

Roche

manufacturer/tradename

Roche

manufacturer/tradename

Roche

manufacturer/tradename

Roche

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

Quality Level

100

Quality Level

100

Quality Level

100

Quality Level

100

General description

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.

Application

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).

Packaging

1 kit containing 6 components.

Quality

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.

Kit Components Only

Product No.
Description

  • Enzyme Mix, Expand High Fidelity 3.5 U/μl

  • PCR DIG Probe Synthesis Mix, containing dATP, dCTP, dGTP (2 mM each) 10x concentrated

  • PCR Buffer with MgCl2 10x concentrated

  • dNTP Stock Solution, containing dATP, dCTP, dGTP, dTTP (2 mM each), pH 7.0 10x concentrated

  • Control Template, plasmid DNA in Tris/EDTA buffer, pH 8.0. The 5-kb plasmid contains the cDNA for the human tissue-type plasminogen activator (tPA) 20 pg/μl

  • Control PCR Primer Mix, containing 50 pmol of each primer, control PCR primer 1 and 2 2 mM each

also commonly purchased with this product

Product No.
Description
Pricing

Hazard Statements

Precautionary Statements

Hazard Classifications

Aquatic Chronic 3

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 2

Flash Point(F)

does not flash

Flash Point(C)

does not flash


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