Recommended Purification and Application for Custom DNA Synthesis
| CARTRIDGE
|
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, hybridization plus gene synthesis, primer extension, squencing, DNA fingerprinting, cycle sequencing, RT-PCR, in situ hybridization, microsatellite polymorphisms.
|
| HPLC |
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,hybridization. Recommended for oligos with < 45 mers.
|
| PAGE |
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
plus gene synthesis and primer extension. Recommended for oligos
with >45 mers. |
|