12039672910

Roche

DIG Northern Starter Kit

greener alternative

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.

shipped in

dry ice

storage temp.

−20°C

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

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).
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.

Packaging

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.
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

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

RIDADR

NONH for all modes of transport

WGK Germany

WGK 2

Flash Point(F)

does not flash

Flash Point(C)

does not flash

Certificate of Analysis

Certificate of Origin

Tuong Vi T Dang et al.
BMC research notes, 7, 655-655 (2014-09-19)
In plants, RNA- based gene silencing mediated by small RNAs functions at the transcriptional or post-transcriptional level to negatively regulate target genes, repetitive sequences, viral RNAs and/or transposon elements. Post-transcriptional gene silencing (PTGS) or the RNA interference (RNAi) approach has...
Interleukin-34 inhibits hepatitis B virus replication in vitro and in vivo
Cheng ST, et al.
PLoS ONE, 12 (2017)
A Novel Virus of the Genus Cilevirus Causing Symptoms Similar to Citrus Leprosis
Roy A, et al.
Phytopathology, 103, 488-500 (2013)
Ziqiang Wang et al.
The Journal of biological chemistry, 291(51), 26377-26387 (2016-10-28)
Once it enters the host cell, herpes simplex virus type 1 (HSV-1) recruits a series of host cell factors to facilitate its life cycle. Here, we demonstrate that serine/arginine-rich splicing factor 2 (SRSF2), which is an important component of the...
Zhihua Wang et al.
Nature medicine, 22(10), 1131-1139 (2016-09-13)
Epigenetic reprogramming is a critical process of pathological gene induction during cardiac hypertrophy and remodeling, but the underlying regulatory mechanisms remain to be elucidated. Here we identified a heart-enriched long noncoding (lnc)RNA, named cardiac-hypertrophy-associated epigenetic regulator (Chaer), which is necessary...
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.
Read More
Protocols
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.
Read More

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