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Custom RNA Oligonucleotide Synthesis

Quality Control Standards

Gene Link oligos are for demanding applications and consistent results. We believe that investigators who value time and have no room for an experiment to fail due to oligo quality should consider Gene Link.

Trityl monitoring coupling efficiency

Our numerous quality control steps for each oligo assure confidence. We maintain an absolute standard of coupling efficiency threshold of greater than 99.5% for all oligos by using the best reagents. Trityl monitoring coupling efficiency of each base added during synthesis with programmed ‘halt’ to seek user intervention if it falls below the threshold. This is not much evident when comparing short oligos but is a requirement for long oligos. Ask our competitors how often they synthesize 200 to 250 mer. Please see the coupling efficiency table and graph down below. Gene Link specializes in long oligos. Our description of long oligos is 180mer to 250mer. You are invited to compare.

Actual Trityl Coupling Efficiency of a 210 mer

Polyacrylamide Gel

Each oligo is run side by side on a polyacrylamide gel to visually assess quality. A real gel picture is included as part of the oligo report.

Crude Oligo Gel Gel Purified Oligo Gel

Crude Oligo Gel Electrophoresis

Polyacrylamide gel electrophoresis of crude oligos. Approximately 8 µg of crude unpurified oligo were loaded to show the truncated failure sequences if any. Lanes 1 ‑ 7: Oligo sizes, 51, 64, 48, 42, 25, 18 and 20.

Results:

A major single band represents high purity of the crude oligonucleotide. The above gel picture shows the quality of Gene Link oligos. All Gene Link oligos are provided with an actual gel picture.

Gel Purified Oligo Gel Electrophoresis

Polyacrylamide gel electrophoresis of crude and gel purified oligos in adjacent lanes. Lanes 1 & 2: 78 mer; lanes 3 & 4: 96 mer; lanes 5 & 6: 161 mer; lanes 7 & 8: 42 mer.

Results:

At Gene Link we recommend gel purification of all long oligos and oligos used in cloning applications. Gel purification is the “gold standard” method of purification as the denaturing polyacrylamide gel resolution approaches single base and the major band is clearly visible to be purified.

Oligo Coupling Efficiency and Expected Yield

PCR and sequencing reactions are very robust and can tolerate up to 50% failure/truncated sequence oligos. However, you are clearly taking a chance by using long oligos synthesized at anything below 99.5% coupling efficiency.
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