- ¥6997.201起订
起订量：
Roche11585606910 DIG-High Prime
- 型号
- Roche11585606910
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详细信息
DIG-High Prime |
| For the nonradioactive labeling of DNA with DIG-11-dUTP alkali-labile, using random oligonucleotides as primers. |
| Cat.No. | Pack Size | Qty | Price | Options |
|---|---|---|---|---|
| 11585606910 | 160 µl for 40 labeling assays 0.01-3 µg template DNA per assay | 6997.2 | ||
Application
DIG-labeled probes are generated at high yield within one hour or after overnight incubation.
The standard reaction with 1 µg template DNA will routinely give a yield of 0.8 µg DIG-labeled DNA after 1 hour, and 2.3 µg after a 20-hour incubation.
labeled DNA probes are used in Southern, northern, and dot blots, as well as in colony or plaque hybridization.
Due to highly specific and sensitive detection systems, DIG-labeled DNA probes can be used for single-copy gene detection in 1 µg total human DNA.
The use of the alkali-labile form of DIG-dUTP, in which the digoxigenin moiety is connected to the spacer arm via an alkali-labile ester bond （Figure 1）, enables easier and more efficient stripping and reprobing of blots.
Benefits
- DNA amounts ranging from 10 ng to 3 µg in a standard reaction.
- DNA of different lengths ranging from small restriction fragments to λ or cosmid DNA.
- DNA, supercoiled or linearized.
- DNA in low melting-point agarose.
Product Description
A standard labeling reaction with 1 µg template yields 0.8 µg newly synthesized digoxigenin-labeled DNA after 1 hour, and 2 µg after a 20-hour incubation at 37°C.
Background Information
, a novel 5x conc. labeling mixture contains optimal concentrations of random primers, nucleotides, and DIG-dUTP, alkali-labile, in a highly efficient reaction buffer with Klenow enzyme. Simply add to the denatured template DNA. The number of pipetting steps are thus reduced to a minimum, while reproducibility and accuracy of experiments are effectively increased.
DIG-labeled probes in the reaction mix or in the hybridization buffer are stable for more than 12 months stored at -15 to -25°C. They can be reused several times if freshly denatured before use.
Figure 1: Structure of alkali-labile DIG-11-dUTP.
Contents
Typical Experiment
Table 1: Yield of labeling reaction.
Quality
In a standard assay with 1 µg linearized pBR 328, 0.8 µg of DIG-labeled DNA is synthesized after 1 hour, and 2.3 µg after 20 hours. When this labeled DNA is used for hybridization at a concentration of 20 ng/ml, 0.03 pg homologous DNA are detected by chemiluminescence with the anti-DIG-alkaline phosphatase conjugate and CSPD on a dot or Southern blot.
