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12039672910

Roche

DIG Northern Starter Kit

greener alternative

suitable for Northern blotting, sufficient for 10 labeling reactions

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Synonym(s):
DIG system, Northern blot

usage

sufficient for 10 blots (blots of 10 x 10 cm2)
sufficient for 10 labeling reactions

Quality Level

manufacturer/tradename

Roche

greener alternative product characteristics

Designing Safer Chemicals
Learn more about the Principles of Green Chemistry.

parameter

68 °C optimum reaction temp.

technique(s)

Northern blotting: suitable

greener alternative category

storage temp.

−20°C

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117458329101158561491011175025910
DIG Northern Starter Kit suitable for Northern blotting, sufficient for 10 labeling reactions

Roche

12039672910

DIG Northern Starter Kit

usage

sufficient for 10 blots (blots of 10 x 10 cm2)

usage

sufficient for 12 labeling reactions, sufficient for 24 blots

usage

sufficient for 12 labeling reactions (10 ng to 3 μg per assay), sufficient for 24 blots (blots of 100 cm2)

usage

sufficient for 2 x 10 labeling reactions

manufacturer/tradename

Roche

manufacturer/tradename

Roche

manufacturer/tradename

Roche

manufacturer/tradename

Roche

greener alternative product characteristics

Designing Safer Chemicals
Learn more about the Principles of Green Chemistry.

greener alternative product characteristics

Designing Safer Chemicals
Learn more about the Principles of Green Chemistry.

greener alternative product characteristics

Designing Safer Chemicals
Learn more about the Principles of Green Chemistry.

greener alternative product characteristics

Designing Safer Chemicals
Learn more about the Principles of Green Chemistry.

parameter

68 °C optimum reaction temp.

parameter

-

parameter

-

parameter

-

greener alternative category

Aligned,

greener alternative category

, Aligned

greener alternative category

Aligned,

greener alternative category

, Aligned

General description

The DIG Northern Starter Kit is designed for the novice DIG system user. It contains the reagents proven to provide successful, reproducible results in northern blots. Additionally, the more convenient forms of standard DIG system products are included (e.g., CDP Star, ready-to-use). The small number of reactions allows the user to gain a firm foundation using the DIG system, then "graduate" to the standard pack sizes. For optimum success, use this kit with the DIG Wash and Block Buffer Set.This kit is used for the generation of single-stranded, DIG-labeled RNA probes and chemiluminescent detection. Labeled probes are synthesized by the in vitro transcription method.
We are 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 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.

Application

DIG northern starter kit has been used in northern blot hybridization to analyse hepatitis B virus (HBV) RNAs, citrus leprosis virus cytoplasmic type 2 (CiLV-C2) RNAs.
The DIG Northern Starter Kit produces DIG-labeled RNA probes that can be used in conjunction with the supplied chemiluminescent detection reagents for northern blotting techniques. Using linearized DNA as a template, SP6, T3, or T7 RNA Polymerases are used to incorporate DIG-11-UTP into the RNA transcript. After labeling, the DIG-labeled probe is immediately ready for use in hybridization. For convenience, DIG Easy Hyb can be used for hybridization. The DIG Easy Hyb granules are easily reconstituted by adding 64 ml DEPC-treated (RNase-free) water directly to the bottle (once reconstituted, DIG Easy Hyb is stable for 1 month at room temperature). After hybridization, the hybridization solution containing the DIG-labeled RNA probe can be stored at -15 to -25 °C for future re-use (up to 1 year).

Packaging

1 kit containing 11 components.
1 kit for up to 10 labeling reactions and detection of 10 blots, blots of 10×10cm2, reactions of μg DNA, yielding approx. 20μg labeled RNA, each
Sensitivity and specificity: Using 1μg of linearized template DNA, the labeling reaction typically yields 20μg of newly synthesized DIG-labeled RNA within 1 hour. The DIG-labeled RNA probe can detect 0.1pg of homologous DNA or RNA in a dot blot. Rare mRNAs can be detected in 0.1μg of total RNA.

Preparation Note

Working solution: Note: Use sterile, RNase-free solutions and equipment
Assay Time: 9 hours 30 minutes
Sample Materials:
  • Linearized plasmid, including the appropriate RNA polymerase promoter sequence (SP6, T3, T7).
  • Specially prepared PCR productWorking solution: Note: Use sterile, RNase-free solutions and equipment.

Note: The length of the region to be transcribed should be in the range of 200 to 1,000 bp. To avoid RNase contamination the DNA must be phenolized.

Other Notes

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

Kit Components Only

Product No.
Description

  • Labeling Mix 5x concentrated

  • Transcription Buffer 5x concentrated

  • SP6 RNA Polymerase 20 U/μl

  • T7 RNA Polymerase 20 U/μl

  • T3 RNA Polymerase 20 U/μl

  • Anti-Digoxigenin-AP antibody, Fab fragments 750 U/ml

  • DNase I, RNase free 10 U/μl

  • CDP-Star ready-to-use

  • Actin RNA Probe, DIG-labeled Antisense Probe, length 588 bases 10 ng/μl

  • DIG Easy Hyb Granules

  • Blocking Solution 10x concentrated

See All (11)

Pictograms

CorrosionExclamation mark

Signal Word

Danger

Hazard Statements

Hazard Classifications

Acute Tox. 4 Oral - Eye Dam. 1 - Skin Irrit. 2

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 2

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 Northern Starter Kit Protocol & Troubleshooting

In general, it is possible to use DNA probes for northern blots (PCR or random-primed DNA probes). It is however important to keep in mind that DNA probes are best detecting abundant messages, such as housekeeping genes.

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