Biotin-dUTP

General protocols for the enzymatic incorporation of biotin-11-dUTP into DNA probes can be found in Section 1 (this chapter). A particularly interesting modification of the typical enzymatic... 

Label 1.0 pg of probe with biotin-11-dUTP by the nick translation reaction... 

General protocols for the enzymatic incorporation of biotin—11-dUTP into DNA... 

Figure 402 Biotin—11-dUTP is perhaps the most popular nucleotide derivative used for enzymatic biotinylation of oligonucleotides. The 11 designation refers to the number of atoms in its spacer arm.

Nick translation kit Biotin-7-dATP or Biotin-11-dUTP. 

Label 1 pg of DNA probe with biotin-11-dUTP or biotin-7-dATP by nick... 

Labeling of probes for in situ hybridization relies on the incorporation of either a radioisotopic dNTP (e.g., dCTP), or of a nonisotopic molecule, such as biotin-7-dAlT or biotin-11-dUTP, by either nick translation or random priming. The site (s) of hybridization can then be seen using autoradiography with isotopic probes, or immunocytochemically if biotin is incorporated into the probe DNA. It is with the latter form of in situ hybridization methodology that this chapter is concerned. 

Hybridization Mix. 55% formamide, 10% dextran sulfate, 1 mg/ml blocking (degraded salmon sperm) DNA, 1 fig/ml of labeled probe (biotin-11-dUTP labeled), IX SSC. 

Label 1 Xg of probe by nick translation using either biotin-11-dUTP or digoxigenin-11-dUTP, following the manufacturer s instructions. 

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