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11636090910

Roche

PCR DIG Probe Synthesis Kit

sufficient for 25 reaction (50 μL final reaction volume)

Synonym(s):

DIG system, probe

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About This Item

UNSPSC Code:
41105500

usage

sufficient for 25 reaction (50 μL final reaction volume)

Quality Level

manufacturer/tradename

Roche

technique(s)

PCR: suitable

storage temp.

−20°C

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

hcodes

Hazard Classifications

Aquatic Chronic 3

Storage Class

12 - Non Combustible Liquids

wgk_germany

WGK 2

flash_point_f

does not flash

flash_point_c

does not flash


Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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N Toplu et al.
Veterinary pathology, 48(3), 576-583 (2010-05-13)
The present study describes the pathologic changes and cellular apoptosis in the central nervous system (CNS) of fetal and neonatal small ruminants infected with border disease virus (BDV), as demonstrated by immunohistochemistry and in situ hybridization. Abortions of ewes and
Takato Sugiyama et al.
Cell reports, 26(12), 3400-3415 (2019-03-21)
18S non-functional rRNA decay (NRD) eliminates non-functional 18S rRNA with deleterious mutations in the decoding center. Dissociation of the non-functional 80S ribosome into 40S and 60S subunits is a prerequisite step for degradation of the non-functional 18S rRNA. However, the mechanisms
Jean-Félix Dallery et al.
Molecular plant pathology, 20(6), 831-842 (2019-03-30)
The role of histone 3 lysine 4 (H3K4) methylation is poorly understood in plant pathogenic fungi. Here, we analysed the function of CclA, a subunit of the COMPASS complex mediating H3K4 methylation, in the brassica anthracnose pathogen Colletotrichum higginsianum. We
Alix Warburton et al.
PLoS genetics, 14(1), e1007179-e1007179 (2018-01-25)
Integration of human papillomavirus (HPV) genomes into cellular chromatin is common in HPV-associated cancers. Integration is random, and each site is unique depending on how and where the virus integrates. We recently showed that tandemly integrated HPV16 could result in
Handeng Liu et al.
Parasitology research, 112(3), 1011-1020 (2012-12-21)
Microsporidia are a group of obligate intracellular parasites of medical and agricultural importance, which can infect almost all animals, including human beings. Using the genome data of Nosema bombycis, four families of miniature inverted-repeat transposable elements (MITEs) in ribosomal DNA

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