Custom Oligo Specifications
Basic Specifications:
- Chemistry: Eurofins MWG Operon uses the robust and well established ß cyanoethyl
phosphoramidite method
- Synthesizers: High performance, fully-automated, fully-documented, and validated
synthesizers for single, massive parallel, and large-scale syntheses
- Solid Supports: Pre-packed, controlled-pore solid supports
- Synthesis Scales Available:
- 10 nmol, 50 nmol, 200 nmol, 1.0 µmol, and 10 µmol
- Large Scale Synthesis: Yields from milligram to multi-gram amounts
- Reagents: Certified high purity, ultra-dry reagents qualified for use in manufacturing
through proprietary processes
- Typical Synthesis Step Coupling Efficiency Yield: 99% and greater
- Maximum Sequence Length: 125mer (please call technical support for extended lengths)
- Deprotection: Proprietary gas-phase deprotection leading to salt-free oligos.
- Purification (optional):
- HPSF® Purity (High Purity Salt Free)
- QuickLC™ Purity
- Ion Exchange and/or Reversed-Phase HPLC Purity
- Polyacrylamide Gel Electrophoresis (PAGE) Purity
- Final Form: (according to customer request)
- Dried material; Total yield or specific amount in nmol or μg units
- Solution; Specific and/or normalized concentration
- Custom-labeled microfuge tube, 96-well, or 384-well plate
Data Sheet
An Oligonucleotide Data Sheet is provided with each order:
____________________________________________________
Customer #
Order #
P.O. #
_____________________________________________
Tube #
Sequence Name
Sequence 5' to 3'
Yield [OD]
Volume [µL]
Concentration [µM]
Yield [nmol]
Yield [µg]
Oligo Length
Molecular Weight [g]
% G/C Content
E260; Oligo Specific Extinction Coefficient
Melting Temperature [Tm]
Synthesis Scale
Oligo Purification Level
Vol of buffer [µl] to add to tube/well to make a 100 µM stock solution
Barcode ID
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Custom Oligo Quality Control (QC) and Quality Assurance (QA)
Quality is an integral part of the Eurofins MWG Operon oligonucleotide synthesis
service. Ordering via the Eurofins MWG Operon website at www.operon.com eliminates
data re-entry errors. To allow order tracking throughout the manufacturing process,
each order and each sequence within that order are automatically assigned unique
numbers.
Eurofins MWG Operon uses proven high quality raw materials from reliable suppliers;
all reagents are lot tested and must pass purity tests prior to use. Fully-automated,
barcode-equipped production systems capture real-time synthesis information for
each oligo, and all oligos are analyzed by UV spectrophotometry to obtain an optical
density (OD) measurement.
All oligos are analyzed by MALDI-TOF or ESI mass spectrometry, depending upon oligo
length. Selected oligos longer than 50 bases are analyzed by polyacrylamide gel
electrophoresis (PAGE) and statistical analyses are performed to monitor oligo quality.
In addition, oligos are regularly sampled for analysis by capillary gel electrophoresis
(CGE).
Any oligo that fails to meet our specifications is resynthesized. Our stringent
quality processes result in high quality oligonucleotides.
The final yield of each oligo is determined by measuring the absorbance at 260 nm
using an automated spectrophotometer. Absorbance readings are transferred into the
Eurofins MWG Operon network database. Conversion of readings to microgram and nanomole
equivalents is computerized.
Bio-QC
To complement the standard processes used to ensure oligo quality, Eurofins MWG
Operon is the only oligonucleotide manufacturer that tests the functionality of
our oligos using a Bio-QC quantitative PCR (qPCR) assay. Performance of the oligo
is determined by attaining the threshold cycle (C
T value) in the qPCR reaction.
C
T is defined as the first PCR cycle with a detectable fluorescence signal over
a defined background level. Oligos synthesized in all of our worldwide manufacturing
facilities are centrally tested at our headquarters in Huntsville, AL, ensuring
that our customers receive oligos of consistently high quality.