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11175025910 Roche

DIG RNA Labeling Kit (SP6/T7) Green Alternative

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Properties

Related Categories 12 Principles Aligned Products, Chemical Synthesis, Greener Alternative Products, Life Sciences
usage   sufficient for 2 x 10 labeling reactions
mfr. no.   Roche
greener alternative product characteristics   Designing Safer Chemicals
Learn more about the Principles of Green Chemistry.
shipped in   dry ice
storage temp.   −20°C

Description

General description

Download the CofA, SDS and IFU specific to your product lot and format

Assay Time: 145minutes
Sample Materials
• Linearized plasmid DNA
• PCR product
Principle
The DIG RNA Labeling Kit produces DIG-labeled, single-stranded RNA probes of known length. Either SP6 or T7 RNA polymerase transcribes these probes in vitro from template DNA (in the presence of digoxigenin-UTP).
RNA Labeling by in vitro Transcription
The DNA to be transcribed is cloned into the polylinker site of appropriate transcription vectors (e.g., pSPT 18 or 19), which contain promoters for SP6 and T7 RNA polymerases. Adjacent template DNA is linearized at a suitable site and the RNA polymerases are used to produce "run off" transcripts. DIG-UTP is incorporated into the transcript. Every 20 to 25th nucleotide of the newly synthesized RNA is a DIG-UTP. Since the nucleotide concentration does not become limiting in the standard transcription reaction, this reaction can generate large amounts of labeled RNA.

Kit for the labeling of RNA with digoxigenin-UTP by in vitro transcription with SP6 and T7 polymerases. By this method, single-stranded RNA probes of known length are produced, which can be used in a variety of hybridization techniques.

Sigma Life Science is committed to bringing you Greener Alternative Products, which adhere to one or more of The 12 Principles of Greener Chemistry. This product is designed as a safer chemical.  The DIG System was established as a sensitive and cost-effective alternative to using radioactivity for the labeling and detection of nucleic acids. There are many available publications that prove the versatility of the DIG System, so use of radio-labeling is no longer the only option for labeling of DNA for hybridization.

Other Notes

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

Packaging

1 kit containing 12 components.

Quality

Test principle: The DNA template to be transcribed is cloned into the polylinker site of appropriate transcription vectors, which contain promoters for SP6 and/or T7 RNA Polymerases. After linearization at a suitable site, RNA is transcribed in the presence of DIG-UTP. Under standard conditions, approximately 10μg of full-length DIG-labeled RNA is transcribed from 1μg template.

Specificity

Sensitivity and Specificity
DIG-labeled RNA probes can detect single-copy genes in as little as 1 μg of mammalian DNA under the following assay conditions: The hybridization mix contains 20 to 100 ng labeled probe/ ml, and the bound probe is detected with anti-DIG-AP and visualized with the chemiluminescent substrate CDP-Star.
Heat inactivation: Stop the reaction by adding 2 μl 0.2 M EDTA (pH 8.0).

Application

For RNA labeling with DIG-11-UTP by in vitro transcription with SP6 and T7 RNA Polymerases. DIG-labeled "run off" transcripts are synthesized with high efficiency and can be used in a variety of hybridization techniques:
• Northern blots
• Southern blots
• In situ hybridizations
• Plaque or colony lifts
• RNase protection experiments
Due to highly specific and sensitive detection systems, DIG-labeled probes can be used for single-copy gene detection in 1μg total human DNA.
Note: Since the linkage between DIG and UTP is resistant to alkali, DIG-labeled RNA can be fragmented by alkaline treatment. Slightly reducing the size of the DIG-labeled RNA probe may make it more suitable for certain applications in in situ hybridization.

Price and Availability


DIG Go Green

Kit component only

Description

Product #

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pSPT18 DNA 0.25 mg/ml SDS    
pSPT19 DNA 0.25 mg/ml SDS    
Control DNA 1, pSPT18-Neo, cleaved with Pvu II 0.25 mg/ml SDS    
Control DNA 2, pSPT19-Neo, cleaved with Pvu II 0.25 mg/ml SDS    
DIG-labeled Control RNA, DIG-labeled "antisense" neo RNA 100 ng/µl SDS    
Unlabeled Control RNA, neo poly (A) "sense" RNA 200 µg/ml SDS    
NTP Labeling Mixture 10x concentrated SDS    
Transcription Buffer 10x concentrated SDS    
DNase I, RNase-free 10 U/µl SDS    
Protector RNase Inhibitor 20 U/µl SDS    
SP6 RNA Polymerase 20 U/µl SDS    
T7 RNA Polymerase 20 U/µl SDS    
Safety & Documentation

Safety Information

Symbol 
GHS07  GHS07
Signal word 
Warning
Hazard statements 
Precautionary statements 
RIDADR 
NONH for all modes of transport

Documents

Certificate of Analysis

Protocols & Articles

Protocols

DIG RNA Labeling Kit (SP6/T7) Protocol & Troubleshooting

Evaluation of DIG RNA labeling efficiency: 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 ...
Keywords: AGE, Digestions, Electrophoresis, Gel electrophoresis, Gene expression, PAGE, Polymerase chain reaction, Precipitation, Purification, Transcription

Peer-Reviewed Papers
15

References

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