I have designed my own oligo probes. How can I determine their specificity?
There are different ways for controlling oligonucleotide specificity, which differ in terms of cost in time and meaningfulness. The simplest and best method is the use of the same material which is not expressing the target. Unfortunately this type of control does not often exist – Usually one or several of the following methods are applied: (a) DNase digestion using probes against DNA, (b) RNase digestion using probes against RNA, (c) oligonucleotides with several mismatches, (d), random oligonucleotides of the same nucleotide composition (e) sense oligonucleotides (RNA targets), (f) reverse oligonucleotides and (g) competition with unlabelled oligonucleotides.
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