Overview

Oligonucleotides produced by GenScript with state-of-the-art oligo-synthesizer have substantial purity with low truncated sequences. There is no further purification for applications involving in Polymerase Chain Reaction(PCR), DNA sequencing and gene synthesis. However oligonucleotides are always contaminated with products of incomplete synthesis and deprotection product, improperly synthesized oligonucleotides, and protecting groups cleaved from the bases after synthesis. The decision as to whether or not purification is required depends on an evaluation of the probable degree of contamination of the desired full-length product (n-mer) with these undesired side products and the possible effects of these contaminants on the intended experiment.

In general, purification steps perform three significant functions

Separation of full-length n-mers from incomplete products (n-1 mer, n-2 mer, et al).
Removal of oligonucleotides resulting from incomplete deprotection, depurination, dimerization, branching, et al.
Removal of cleaved blocking groups from bases.

Purification Options

Oligonucleotides purification can be accomplished by various methods, The purification method selected depends on the size of the oligonucleotide, the degree of purity required with the application. The common techniques available and used frequently are polyacrylamide gel purification (PAGE), ion exchange (IE) or reverse phase (RP) HPLC, and reverse phase cartridges such as the RPC. The table below summarizes the purification range of each of the above techniques.

Length RPC PAGE HPLC
Ion Exchange Reverse phase
<15 mer
No
No
Yes
Yes
15-40 mer
Yes
Yes
Yes
Yes
41-60 mer
Yes
Yes
Yes
No
60~120 mer
No
Yes
No
No

Purification Options

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