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
Related Categories
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.
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.
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.
Signal Word
Warning
Hazard Statements
Precautionary 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 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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