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DIG Oligo Tailing Kit, 2nd gen
Contents
Reaction Buffer, 5x, 100 µl
CoCl2 Solution, 100 µl, 25 mM
DIG-dUTP Solution, 25 µl, 1 mM
dATP Solution, 25 µl, 10 mM
Recombinant Terminal Transferase, 25 µl, [400 U/µl]
Control Oligonucleotide, unlabeled, 25 µl, [20 pmol/µl]
Oligonucleotide, DIG-dUTP/dATP tailed, 20 µl, [2.5 pmol/µl]
Control DNA, 20 µl, [0.25 mg/ml]
Glycogen Solution, 50 µl, [20 mg/ml]
DNA Dilution Buffer, 3 x 1 ml, [50 µg/ml] fish sperm DNA
poly(A) Solution, 1 ml, [10 mg/ml]
Application
Oligonucleotides, tailed with DIG-dUTP, are used for DNA or RNA hybridization in membrane blots or in situ hybridization.
Benefits
Very sensitive hybridization probes, due to the incorporation of several DIG-nucleotides
Fast hybridization kinetics, due to the small size of oligonucleotides
Single-stranded probes, no renaturation during hybridization
Sequence can be designed according to the experiment
Specially suited for in situ hybridization; due to their small size, oligonucleotides readily diffuse into fixed tissues and cells
Product Description
Tailing of oligonucleotides at the 3'-end with DIG-11-dUTP and recombinant Terminal Transferase. Oligonucleotides are tailed with DIG-dUTP and dATP at an average tail length of 50 nucleotides (tail length range: 10 – 100).
Principle
DIG-dUTP and dATP are combined at a concentration that gives the highest DIG incorporation into the tail, and optimal spacing of DIG and dATP, to achieve the highest sensitivity in hybridization experiments. DIG-dUTP and dATP are provided as separate solutions to allow greater flexibility in terms of tail length, hapten spacing, and the use of unlabeled nucleotide(s).
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