Custom DNA & qPCR Probes
Standard custom from Bioneer
For routine applications, Bioneer offers standard custom oligos with no modification, Bio-RP purification, provided in a tube format.
Standard oligonucleotide synthesis service includes synthesis of up to 130-mer oligonucleotides. A variety of synthesis scales, as well as a choice of purification methods, (Bio-RP, HPLC or PAGE), depending on your application, can be selected.
(Guaranteed quantity of starting materials)
|25 nmol, 50 nmol, 0.2 umol, 1.0 umol, 10umol and 15 umol|
|Length of oligomer||5-130 mer|
|Available purification methods||Bio-RP, HPLC and PAGE|
|Q.C & Q.A||MALDI-TOF Mass Spectrometer & CE (Capillary Electrophoresis)|
Synthesis Scale represents the moles of starting material. Final yield is determined by the following factors: Under the assumption that coupling efficiency of synthesis is 99 %, 98 %, or 97 % respectively, final yield of 30-mer should be 0.99 x 30 = 74 %, 0.98 x 30 = 55 %, and 0.97 x 30 =40 %. Thus, the longer oligo is, the less final yield becomes. Coupling efficiency is dependent on several factors including the sequence of the oligo. In fact, it is possible for an oligo with a specific sequence not to be synthesized. Purification methods also give differences in final yield. Desalted oligo shows higher final yield, due to presence of significant amounts of uncoupled n-1, n-2, n-3, … mer oligo. Bioneer's Bio-RP purification process, along with MALDI-TOF QC, isolates pure target oligo of exact length, without n-1 mer impurities.
Various purification methods of Bio-RP, HPLC, and PAGE.
Free Bio-RP purification service only by Bioneer!
Phosphoramidite, which serves as the coupling monomer in oligonucleotide synthesis, has a DMT-protected 5'-OH group to prevent repetitive coupling reaction during the individual addition steps of synthesis. Only the n-mer oligos with the expected length, not n-1, n-2, n-3 … mer oligos, have the 5'-DMT group provided DMT was not removed after final round of synthesis (AKA "Trityl-On" mode). Bioneer developed our unique Bio-RP resin, which attracts the hydrophobic DMT group, as well as an automated column purification process. We can purify n-mer oligo with high purity (>85%) using Bio-RP's efficient isolation of n-mer and removal of n-1, n-2, n-3…mer.
High-purity oligonucleotide is required for applications such as Site-directed mutagenesis or quantitative gene expression experiments. Oligo purification through desalting column or Bio-RP resin is not sufficient for those applications. HPLC purification provides oligo with significantly higher purity. Bioneer uses either anion-exchange resin or reverse-phase (RP) resin for the HPLC purification. Both Anion-exchange chromatography and RP-HPLC purification provide Oligos that are > 90 % pure.. It should be noted that oligos over 35-mer may have lower purity, because longer oligos reduce purification efficiency.
We use PAGE purification method to purify longer oligos with high purity. Electrophoresis on a polyacrylamide gel can separate oligos up to 130-mer, and gives > 95 % purity after the process. PAGE purification has very high separation ability but expect lower final yields because of loss during the extraction process from the gel.
Recommended Purification Methods by Application
|Standard PCR / RT -PCR||√|
|Real Time PCR||√|
|Cloning (e.g. chemical linkers)||√||√|
|DNA Microarray Probes||√||√|
|cDNA library generation||√|
|End labelling & FISH||√|
|Northern & Southern blotting||√||√|
|Gel Shift Assay||√||√|
Yield of oligo synthesis is dependent upon composition of bases, purification method, oligo length, synthesis scale and
the presence of modification(s).
Bioneer guarantees "minimum yield" of an oligo to customers.
|Synthesis Scales||Lowest Guaranteed O.D (for 25-mer)|
Generally DMT-monitoring QC, PAGE QC, or HPLC QC is used for quality control of synthesized oligo,
but its indirect nature does not provide the exact value of failed oligo amount or of successful modification.
Unlike other methods, MALDI-TOF (Matrix-Assisted Laser Desorption / Ionization Time of Flight) mass spectrometer can
get definite quality of synthesized oligo from the measurement of exact molecular weight of oligo,.
Bioneer provides high-quality oligos through MALDI-TOF QC of all products.
QC Data sheet from MALDI-TOF is also provided for confirmation of oligo molecular weight.
HPCE (High Performance Capillary Electrophoresis) 'Oligo-pro™ is used to analyze purity of oligos over 50-merin lenght.
All ordered oligos, except PAGE-purified or modified ones, will be shipped from Korea within 2 days of ordering.
(Note that the Delivery of the final product is subject to delay, in the event of re-synthesis or shipping company's
Delivery to customers site, will vary by country. Please contact your local distributor for a schedule.