Easiest way to Detect Oligonucleotides without Instrumentation
Without the need to have HPLC equipment, the “UV shadowing” method to detect oligonucleotides have been used for over 30 years. The method simply requires TLC (thin layer chromatography) plates with uv indicator; this can be acquired from Whatman cat 4420222) 250um layer, 20x20cm The plate is covered with saran wrap plastic( to keep from getting wet) and the PAGE gel (after the electrophoresis run is completed) is placed on top.Then using a handheld UV lamp (Fotodyne) with short wavelength lamps (250-300nm), uv light is shined on the gel; one clearly can observe the oligonucleotides as dark blue bands. This method requires at least 0.5 OD units of the oligo to be visualized; generally this is not a problem as yields from oligo synthesis, even in the smaller scales, render over 5 OD’s units of product. The UV shadowing method can be used as analytical (0.5-1mm thickness combs) or as a preparative method (3-5 mm thickness combs).
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