DIG 3'-End Oligonucleotide Labeling Kit with Digoxigenin-11-ddUTP

DIG 3'-End Oligonucleotide Labeling Kit with Digoxigenin-11-ddUTP contains all reagents (except oligonucleotide template to be labeled and materials for Digoxigenin detection) required for efficient 3'-End digoxigenylation of DNA oligonucleotides (length: 20-100 bp, 5 pmol per reaction) . The labeling principle is based on Terminal deoxynucleotidyl Transferase (TdT) that template-independently transfers 1 Digoxigenin-11-ddUTP to the 3'-OH group of ssDNA (e.g. an oligonu-cleotide) in the presence of CoCl 2. It is similar to the principle of DIG Oligonucleotide 3'-End DNA Labeling Kit, 2nd Generation (Roche) . The resulting 3'-End digoxigenylated oligonucleotides are ide-ally suited for applications involving sequence-specific protein binding or hybridiziation such as EMSA, Northern or Southern blots. Compared to internal, random digoxigenylated probes, Digoxigenin is located at the 3'-End only and less likely interferes with probe binding. TdT possesses a preference for single-stranded DNA (ssDNA) over dsDNA with 3'-overhangs or blunt ends. For the preparation of labeled dsDNA complexes, label each complementary oligonucleotide separately and anneal them before use.