Custom Oligo Purification Options
Salt Free Purity
Salt-Free is our basic oligo purity option and is appropriate for most unmodified
oligos used in many applications, including routine PCR, microarray fabrication,
and blotting.
HPSF® Purity
High purity salt free (HPSF) purification is a high-performance, high-throughput
purification that efficiently removes most N-1 and other truncated sequences created
during oligo synthesis. When using unmodified oligos in more sensitive applications
such as qPCR or DNA sequencing, HPSF purification delivers excellent yields and
improved purity.
HPSF purity is available and recommended for oligos containing:
- IUB Wobbles (degenerate nucleotide sites)
- 5' Amino C6 Linker [AminoC6]
- 5' Phosphate [Phos]
- 5' Biotin-TEG [BioTEG]
QuickLC™ Purity
QuickLC purification: Twice the Yield in Half the Time of HPLC; Same Purity—Guaranteed!
QuickLC purification is a proprietary purification process that offers purity levels
of 85% and above*—equal to or greater than the levels obtained by HPLC purification.
Extensive testing in end user laboratories has proven that oligos purified with
the QuickLC method perform identically to oligos that go through the older HPLC
process. If you believe that your QuickLC purity oligos are not performing as good
or better than HPLC-purified oligos, call us and we’ll make it right.
The QuickLC process removes most N-1 and other truncated failure sequences created
during synthesis and achieves purity levels of 85% and above*. Selection of QuickLC
purification ensures delivery of your oligo at HPLC-grade purity; typically within
2 days!
QuickLC is highly recommended for oligos needing HPLC-grade purity or when higher
than typical HPLC yields are required for your experiment. For a list of oligos
that can be purified using our QuickLC method, see the Yield Guarantees page of
our website.
QuickLC purity is available and recommended for oligos containing:
- IUB Wobbles (degenerate nucleotide sites)
- 5' 6-FAM [6-FAM]
- 5' Amino C6 Linker [AminoC6]
- 5' Phosphate [Phos]
- 5' Biotin-TEG [BioTEG]
HPLC Purity
Standard high performance liquid chromatography (HPLC) purification is recommended
for oligos that require additional purification, such as dual-labeled qPCR probes,
Molecular Beacons, and most modified oligos that cannot be purified by our QuickLC
purification method.
HPLC purification removes most N-1 and other truncated failure sequences created
during synthesis and achieves purity levels of 85% and above*. Oligos purified by
HPLC typically ship within 5 days. When faster delivery times are critical, Eurofins
MWG Operon encourages you to consider the QuickLC purification, where it is offered
for you oligo type.
Our oligo purification experts will determine and employ the optimum HPLC technique
for your specific oligos and select only the fractions containing the highest purity
for delivery to your lab. The techniques employed include (1) reversed-phase HPLC
purification that separates oligos on the basis of hydrophobicity; (2) ion exchange
HPLC that separates oligos on the basis of charge. 2-step chromatographic purification
(reversed phase followed by ion exchange) is recommended for oligos longer than
70 bases or for oligos used in applications that require the highest HPLC purity
and is detailed below.
PAGE Purity
Polyacrylamide gel electrophoresis (PAGE) purification is recommended for oligos
that require the highest level of purification, and achieves purity levels for unmodified
oligos of 90% and above
*. Although PAGE purification achieves the highest purity
levels available, recovery of the oligo from the matrix is an inefficient process,
resulting in lower final yields. Thus PAGE purification is not recommended as a
routine technique. PAGE purification also cannot be used for oligos shorter than
20 bases.
PAGE separates oligos on the basis of molecular weight and is performed on a denaturing
polyacrylamide gel prepared with stock solutions of acrylamide, urea, and TBE buffer.
The desired oligo band is detected through UV shadowing and the full-length product
is excised and eluted from the gel.
2-Step Purity – Reversed-Phase/Ion Exchange
This 2-step purification process uses reversed-phase chromatography followed by
anion exchange purification, allowing a two-dimensional purification based on both
hydrophobicity and charge. This purity level is highly recommended for all oligos
70 bases or longer.
2-Step Purity – Reversed-Phase/PAGE
This 2-step purification process uses reversed-phase (RP) chromatography followed
by PAGE, allowing a two-dimensional purification based on hydrophobicity and molecular
weight. This purification level is highly recommended for oligos 70 bases or longer
that are used in applications that require PAGE purification. 2-step RP/PAGE purification
is also recommended for oligos used in applications that require the highest PAGE
purity, such as crystallography and gel-shift assays.
Recommended purification method by application
|
Application |
Oligo |
Salt-Free |
HPSF® |
QuickLC™ |
HPLC |
PAGE |
|
Standard PCR* and qPCR |
Unmodified |
X |
X |
|
|
|
|
Multiplex PCR |
Unmodified |
|
X |
X |
X |
|
|
Sequencing |
Unmodified |
|
X |
|
|
|
|
Microarray |
Unmodified |
X |
X |
|
|
|
|
Modified |
|
X |
X |
X |
|
|
Blotting |
Unmodified |
X |
X |
|
|
|
|
Modified |
|
X |
X |
X |
|
|
cDNA Library |
Unmodified |
X |
X |
|
|
|
|
Construction |
|
|
|
|
|
|
|
Primer Extension |
Unmodified |
|
X |
|
|
|
|
Modified |
|
|
X |
X |
|
|
454 Fusion Primer |
Unmodified |
|
X |
X |
X |
|
|
Modified |
|
|
|
X |
|
|
Antisense |
Modified |
|
X |
X |
X |
|
|
Cloning (Chemical Linkers) |
Unmodified |
|
X |
X |
X |
X |
|
End Labeling and FISH |
Modified |
|
|
|
X |
|
|
Genotyping / SNPs |
Unmodified |
|
X |
X |
X |
|
|
Modified |
|
|
|
X |
|
|
Kinasing |
Modified |
|
|
|
X |
|
|
Primers with Restriction Sites |
Unmodified |
|
X |
X |
X |
|
|
qPCR |
Modified |
|
|
X |
X |
|
|
Crystallography |
Unmodified |
|
|
|
|
X |
|
Gel Shift Assays |
Unmodified |
|
|
|
|
X |
|
Gene Synthesis |
Unmodified |
|
|
|
|
X |
|
Mutagenesis |
Unmodified |
|
|
|
|
X |
These are general guidelines for common applications, but customers are free to
choose for themselves the appropriate purification. Available purification options
are determined by oligo length, modifications, and synthesis scale.
HPSF and QuickLC are trademarks of Eurofins MWG Operon.
*We cannot guarantee the purity of oligos longer than 70 bases. For oligos longer than 70 bases, Eurofins MWG Operon strongly recommends one of our 2-Step Purity options.