英文名称： DIG Nothern Starter KitROCHE
包含标记DIG RNA及进行Northern印迹的所有必需试剂, 配合DIG Wash and Block Buffer Set方便进行Northern印迹; 能作10个标记反应及10次检测(100mm2膜)。特别适合初用者使用; 标记混合物经最优化, 配合提供的化学发光底物CDP-Star, 能产生最高的灵敏度。
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).
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 g of linearized template DNA, the labeling reaction typically yields 20 g 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 g of total RNA.
Background Information
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.
Contents
- Labeling Mix, 40 l, 5x conc.
- Transcription Buffer, 40 l, 5x conc.
- SP6 RNA Polymerase, 20 l, [20 U/l]
- T7 RNA Polymerase, 20 l, [20 U/l]
- T3 RNA Polymerase, 20 l, [20 U/l]
- Anti-digoxigenin-AP, Fab fragments, 60 l, [750 U/ml]
- DNase I, RNase free, 20 l, [10 U/l]
- CDP-Star, ready-to-use, 20 ml
- Actin RNA Probe, DIG-labeled Antisense Probe, length 588 bases, [10 ng/l]
- DIG Easy Hyb Granules, 2 bottles for 100 ml solution each
- Blocking Solution, 2 x 100 ml, 10x conc.