DIG Easy Hyb
英文名称: DIG Easy Hyb
型号:null    产品货号: 11603558001
价格:请致电:010-57128832,18610462672
品牌: roche

DIG Easy Hyb can be used in all membrane hybridization applications using nonradioactive (e.g., digoxigenin-labeled) nucleic-acid probes. DIG Easy Hyb is ready made, and can be used without any additions. It is specially designed for use with DIG kits.

Non-toxic solution, does not contain formamide.

Absence of contaminants: DNase- and RNase-free.

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DIG-High Prime 高效地高辛DNA标记试剂,标记DNA探针, 只需一步操作, 标记效率较通常随机引物法高3倍。40 T

Application

DIG-High Prime is used for the highly efficient random-primed labeling of DNA probes with DIG-11-dUTP, alkali-labile.
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.
DIG-High Prime 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

DIG-High Prime guarantees efficient labeling of:

Product Description

Labeling efficiency: 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

DIG-High Prime, 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 DIG-High Prime 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

5x conc. labeling mixture, 160 µl containing random primer mixture, 1 U/µl Klenow enzyme, labeling grade, 1 mM dATP, 1 mM dCTP, 1 mM dGTP, 0.65 mM dTTP, 0.35 mM DIG-11-dUTP, alkali-labile, and 5x stabilized reaction buffer in glycerol, 50 (v/v).

Typical Experiment



Table 1: Yield of DIG-High Prime labeling reaction.

Quality

Function test: 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.

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