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DIG Northern Starter Kit
For transcription-labeling of RNA with digoxigenin and SP6/T7/T3 RNA polymerases and chemiluminescent detection with CDP-Star, ready-to-use.
Contents
1.Labeling Mix, 40 µl, 5x conc.
2.Transcription Buffer, 40 µl, 5x conc.
3.SP6 RNA Polymerase, 20 µl, [20 U/µl]
4.T7 RNA Polymerase, 20 µl, [20 U/µl]
5.T3 RNA Polymerase, 20 µl, [20 U/µl]
6.Anti-digoxigenin-AP, Fab fragments, 60 µl, [750 U/ml]
7.DNase I, RNase free, 20 µl, [10 U/µl]
8.CDP-Star, ready-to-use, 20 ml
9.Actin RNA Probe, DIG-labeled Antisense Probe, length 588 bases, [10 ng/µl]
10.DIG Easy Hyb Granules, 2 bottles for 100 ml solution each
11.Blocking Solution, 2 x 100 ml, 10x conc.
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).
Product Description
Sensitivity and specificity: Using 1 µg of linearized template DNA, the labeling reaction typically yields 20 µg of newly synthesized DIG-labeled RNA within 1 h. The DIG-labeled RNA probe can detect 0.1 pg of homologous DNA or RNA in a dot blot. Rare mRNAs can be detected in 0.1 µg of total RNA. |