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).

General description

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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.

The DIG Northern Starter Kit was 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.The 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.

Other Notes

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

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.