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CARTRIDGE
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Typical yield of
Reverse-phase cartridge purification is 80-90%.
The basis of the separation is the difference in hydrophobicity between full length product (which contains a 5'-DMT) and truncated sequences (without DMT groups). Because the differences in hydrophobicity between the full length-DMT product and non-DMT truncated sequences are reduced as the oligo length is increased, cartridge purification is NOT recommended for oligos > 45 bases.
Applications: PCR, probing antisense, mobility shift, arbitrary 10-mer applications, |
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HPLC
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Typical HPLC product yields is 90-97% purity. The capacity and resolving properties of HPLC columns are also much greater than cartridge devices, so HPLC is the method of choice for purifying larger quantities of oligos (i.e. >= 1 umol). As with cartridges, reverse-phase HPLC is usually NOT recommended for purifying oligos longer than 45 bases
Applications:
PCR, probing antisense, mobility shift, arbitrary 10-mer applications, |
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PAGE
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The basis of the separation is charge and MW. In most cases , full length (n) oligo can be separated from oligos only one base shorter (n-1) with excellent resolution of 95-99% purity can be achieved. ). PAGE is the recommended technique for purifying oligos >45 bases long. Yields from PAGE are lower than from other methods due to the relative inefficient extraction of oligos from the gel.
Applications: PCR, probing antisense, mobility shift, arbitrary 10-mer applications |
