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11277073910

Roche

DIG RNA Labeling Mix

sufficient for 20 reactions, solution

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Synonym(s):
nucleic acid labeling

form

solution

Quality Level

usage

sufficient for 20 reactions

packaging

pkg of 40 μL

manufacturer/tradename

Roche

storage temp.

−20°C

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DIG RNA Labeling Mix sufficient for 20 reactions, solution

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packaging

pkg of 40 μL

packaging

pkg of 50 μL

packaging

pkg of 500 μL (2 x 250 μl)

packaging

pkg of 160 μL

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

usage

sufficient for 20 reactions

usage

sufficient for 25 standard reactions

usage

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

usage

sufficient for 40 labeling reactions

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


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Articles

Digoxigenin (DIG) Labeling Methods

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

DIG RNA Labeling Mix Protocol Troubleshooting

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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