Labeling rna oligos DNA polymerase Labelling Oligonucleotides Annealing Complementary Oligo


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labeling rna oligos

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	Radioactive Oligo labelling of DNA
	The procedure is based on the Klenow fragment, or large subunit, of DNA polymerase I. Klewnow retains the 5' - 3' polymerase activity but lacks the 5' - 3' exonuclease activity. Since Klewnow can not begin synthesis without a primer, a synthetic set of primers (6 bp random oligomers) are added to the single stranded DNA template.

	Labelling Oligonucleotides
	Mix equal volumes of both complementary oligos (at equimolar concentration) in a 1.5ml microfuge tube. Place tube in a standard heatblock at 90 - 95ºC. Remove the heatblock from the apparatus and allow to cool to room temperature (or at least below 30ºC) on the workbench.

	Annealing Complementary Oligo
	A simple and cheap way to make a short (< 100 bp) piece of DNA is to order two complementary primers from a company such as Invitrogen. Other options are described under part fabrication and annealing and primer extension. Having annealed the primers, you most likely want to use them as an insert into a vector (the linked page describes BioBrick vectors).