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11277073910

Roche

DIG RNA Labeling Mix

sufficient for 20 reactions, solution

Synonym(s):

nucleic acid labeling

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About This Item

UNSPSC Code:
41105500

form

solution

Quality Level

usage

sufficient for 20 reactions

packaging

pkg of 40 μL

manufacturer/tradename

Roche

impurities

Ribonuclease, none detected (up to 20 µl using MSII-RNA)

color

colorless

solubility

water: miscible

storage temp.

−20°C

General description

Labeling efficiency: Approximately 10μg of full-length digoxigenin-labeled RNA is transcribed from 1μg linear template DNA.
Assay Time: 135 minutes
Sample Materials
Linearized plasmid DNA:
The DNA to be transcribed is cloned into the polylinker site of an appropriate transcription vector which contains adjacent to the polylinker a promoter for SP6, T7 or T3 RNA polymerase. For the synthesis of ‘run off′ transcripts the plasmid is linearized by a restriction enzyme. Restriction enzymes creating 5′-overhangs should be used; 3′ overhangs should be avoided. The linearized template DNA should be purified by phenol chloroform extraction and ethanol precipitation, to avoid RNase contamination. For ′run around′ transcription circular plasmid DNA is used.
PCR product:
PCR-fragments which contain RNA polymerase promoter sequences can also be used as templates for transcription. Purification of the PCR fragment by HighPure column purification prior to transcription is recommended.
DIG-labeled, single-stranded RNA probes of defined length are generated by in vitro transcription. DIG-11-UTP is incorporated by SP6, T7, and T3 RNA polymerases at approximately every 20 to 25th nucleotide of the transcript under standard conditions. The DIG RNA Labeling Mix is specifically designed for the use with SP6, T7, and T3 RNA polymerases, which are supplied with an optimized transcription buffer.
Convenient nucleotide mixture for the labeling of RNA with Digoxigenin-11-UTP.

Contents
10x solution with:
10 mM ATP, CTP, GTP (each), 6.5 mM UTP, 3.5 mM DIG-11-UTP.

Specificity

Heat inactivation: Stop the reaction by adding 2 μl 0.2 M EDTA (pH 8.0).

Application

RNA labeling with Digoxigenin-11-UTP by in vitro transcription with SP6, T7, and T3 RNA polymerases. DIG-labeled RNA is used in a variety of hybridization techniques:
  • Northern blots
  • Southern blots
  • Dot blots
  • Plaque or colony lifts
  • RNase protection experiments
  • Chromosomes, cells, and tissue sections in situ

Features and Benefits

The DIG RNA Labeling Mix is especially designed for the use with SP6,T7 and T3 RNA polymerases from Roche which are supplied with an optimized transcription buffer.

Quality

Function tested in the DIG RNA Labeling Kit and in the DIG Nucleic Acid Detection Kit.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Pictograms

Exclamation mark

Signal Word

Warning

Hazard Statements

Hazard Classifications

Acute Tox. 4 Oral

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

does not flash

Flash Point(C)

does not flash


Certificates of Analysis (COA)

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Jiabin Chen et al.
Cerebral cortex (New York, N.Y. : 1991), 20(3), 650-660 (2009-07-03)
Experience-dependent plasticity of the adult visual cortex underlies perceptual learning and recovery of function following central nervous system lesions. To reveal the signal transduction cascades involved in adult cortical plasticity, we utilized a model of remapping of cortical topography following
Marco Nousch et al.
Journal of cell science, 126(Pt 18), 4274-4285 (2013-07-12)
Post-transcriptional regulatory mechanisms are widely used to control gene expression programs of tissue development and physiology. Controlled 3' poly(A) tail-length changes of mRNAs provide a mechanistic basis of such regulation, affecting mRNA stability and translational competence. Deadenylases are a conserved
Ya Zhang et al.
Open biology, 10(9), 200172-200172 (2020-09-09)
Somatostatin (SS) and allatostatin-C (ASTC) are structurally and evolutionarily related neuropeptides that act as inhibitory regulators of physiological processes in mammals and insects, respectively. Here, we report the first molecular and functional characterization of SS/ASTC-type signalling in a deuterostome invertebrate-the
Cheryl C Hsia et al.
Evolution & development, 12(2), 131-143 (2010-05-04)
We tested whether Artemia abd-A could repress limbs in Drosophila embryos, and found that although abd-A transcripts were produced, ABD-A protein was not. Similarly, developing Artemia epidermal cells showed expression of abd-A transcripts without accumulation of ABD-A protein. This finding
Xiaolin Tian et al.
Molecular and cellular biology, 30(5), 1269-1284 (2009-12-30)
Little is known about how differentiating cells reorganize their cellular structure to perform specialized physiological functions. MIST1, an evolutionarily conserved transcription factor, is required for the formation of large, specialized secretory vesicles in gastric zymogenic (chief) cells (ZCs) as they

Articles

Digoxigenin (DIG) labeling methods and kits for DNA and RNA DIG probes, random primed DNA labeling, nick translation labeling, 5’ and 3’ oligonucleotide end-labeling.

Protocols

Determine the labeling efficiency in terms of μg (expected yield of a standard labeling reaction is 20 μg of DIG labeled RNA per μg linearized template DNA after the DIG RNA labeling reaction).

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