Roche DIG DNA Labeling and Detection Kit
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产品英文名称:DIG DNA Labeling and Detection Kit
产品中文名称:地高辛标记DNA和检测试剂盒
罗氏 DIG DNA 地高辛标记DNA和检测试剂盒的应用:
Random-primed labeling of DNA probes with DIG-11-dUTP, alkali-labile and color detection of DIG-labeled hybrids. DIG-labeled probes are used for hybridization in Southern, northern, and dot blots, as well as for colony and plaque screening.
DIG-labeled hybrids are detected with an anti-DIG-alkaline phosphatase conjugate and the substrates NBT (nitroblue tetrazolium salt) and BCIP (5-bromo-4-chloro-3-indolyl phosphate, toluidinium salt), which give a light-blue precipitate.
Roche DIG DNA Labeling and Detection Kit 组分:
Unlabeled Control DNA 1, 100 µg/ml
2. Unlabeled Control DNA 2, 200 µg/ml
3. DNA Dilution Buffer
4. DIG-labeled Control DNA, 5.2 µg/ml
5. 10x Hexanucleotide Mix
6. 10x dNTP Labeling Mixture
7. Klenow Enzyme, Labeling grade, 2 U/µl
8. Anti-digoxigenin-AP-conjugate, 750 U/ml
9. NBT/BCIP Concentrated Stock Solution
10. Blocking Reagent
Roche 罗氏 DIG DNA 地高辛标记DNA和检测试剂盒 Labeling and Detection Kit 检测原理:
高效DNA地高辛标记检测试剂盒I是用地高辛(Digoxigenin,简称DIG;又称异羟基洋地黄毒甙元,是一种类固醇半抗原分子)标记的DNA探针,应用于分子杂交和随后的显色反应检测。罗氏公司的该试剂盒检测主要分三阶段进行:1)DNA标记:根据随机引物标记技术,生成DIG地高辛标记的DNA探针。2)杂交:按照标准杂交方法进行,将地高辛标记的DNA探针用于核酸点杂交,可选择带正电的尼龙膜或纯硝酸纤维素膜作为杂交膜。 3)免疫学检测:首先将标记有AP的地高辛抗体与杂交探针免疫结合;通过检测碱性磷酸酶与NBT/BCIP底物进行显色反应的数值,从而达到免疫检测的目的。
Roche 罗氏 DIG DNA 地高辛标记DNA和检测试剂盒 Labeling and Detection Kit 实验图像:
罗氏 DIG DNA 地高辛标记DNA和检测试剂盒的产品描述:
Sensitivity and specificity: The standard labeling reaction with 1 µg template will give a yield of 260 ng DIG-labeled DNA after a 1-hour incubation at +37°C, or 780 ng after a 20-hour incubation. The sensitivity of DIG detection depends on the concentration of the DIG-labeled probe in the hybridization reaction, and the duration of the color reaction.
Table 1: Sensitivity of DIG detection.
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