Roche DIG DNA Labeling and Detection Kit
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產(chǎn)品英文名稱:DIG DNA Labeling and Detection Kit
產(chǎn)品中文名稱:地高辛標(biāo)記DNA和檢測試劑盒
羅氏 DIG DNA 地高辛標(biāo)記DNA和檢測試劑盒的應(yīng)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 組分:
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 羅氏 DIG DNA 地高辛標(biāo)記DNA和檢測試劑盒 Labeling and Detection Kit 檢測原理:
高效DNA地高辛標(biāo)記檢測試劑盒I是用地高辛(Digoxigenin,簡稱DIG;又稱異羥基洋地黃毒甙元,是一種類固醇半抗原分子)標(biāo)記的DNA探針,應(yīng)用于分子雜交和隨后的顯色反應(yīng)檢測。羅氏公司的該試劑盒檢測主要分三階段進(jìn)行:1)DNA標(biāo)記:根據(jù)隨機(jī)引物標(biāo)記技術(shù),生成DIG地高辛標(biāo)記的DNA探針。2)雜交:按照標(biāo)準(zhǔn)雜交方法進(jìn)行,將地高辛標(biāo)記的DNA探針用于核酸點(diǎn)雜交,可選擇帶正電的尼龍膜或純硝酸纖維素膜作為雜交膜。 3)免疫學(xué)檢測:首先將標(biāo)記有AP的地高辛抗體與雜交探針免疫結(jié)合;通過檢測堿性磷酸酶與NBT/BCIP底物進(jìn)行顯色反應(yīng)的數(shù)值,從而達(dá)到免疫檢測的目的。
Roche 羅氏 DIG DNA 地高辛標(biāo)記DNA和檢測試劑盒 Labeling and Detection Kit 實(shí)驗(yàn)圖像:
羅氏 DIG DNA 地高辛標(biāo)記DNA和檢測試劑盒的產(chǎn)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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