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DIG DNA Labeling and Detection Kit
For random-primed DNA labeling with digoxigenin-dUTP, alkali-labile and detection of hybrids by enzyme immunoassay.
Contents
1.Unlabeled Control DNA 1, 20 µl, 100 µg/ml
2.Unlabeled Control DNA 2, 20 µl, 200 µg/ml
3.DNA Dilution Buffer, 2 x 1 ml
4.DIG-labeled Control DNA, 50 µl, 5.2 µg/ml
5.10x Hexanucleotide Mix, 50 µl
6.10x dTP Labeling Mixture, 50 µl
7.Klenow Enzyme, labeling grade, 25 µl, 2 U/µl
8.Anti-digoxigenin-AP-conjugate, 200 µl, 750 U/ml
9.NBT/BCIP Concentrated Stock Solution, 10 x 1 ml
10.Blocking Reagent, 2 x 50 g
Application
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.
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