inqaba biotec has the laboratory facilities and technical expertise to meet your demands in the production of high quality synthetic oligonucleotides. We have been in the business of synthesizing custom oligos for more than 6 years

Our labs are proudly South African, allowing us to provide our customers with affordable DNA synthesis options, personal customer consultation and fastest delivery time in South Africa. You can order with confidence with our 100% satisfaction guarantee.

Tabulated below are the different synthesis scales we offer with the respective minimum yield.

* Mass yield is dependent on length of the oligo when measured with optical density

All desalted custom synthesized oligos are sent lyophilized. Please ensure that you spin the primer tube for a minute to ensure that your primer pellet does not escape when opening your tube to dissolve your primer

Minimum Guaranteed yield for standard 20mer oligo: The scale of the oligo synthesis describes the amount of the CPG material to start the synthesis. The yield of the final oligo product is measured at 260nm. 1 OD for a standard 20mer oligo corresponds to approx. 5 nmole of the oligo product

All standard desalted oligos are deprotected and desalted to remove impurities.

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inqaba biotec offers a range of oligonucleotide modifications for both 5' and 3' ends, as well as internal modifications.

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inqaba biotec not only synthesizes primers for conventional PCR, but also probes for quantitative or real-time PCR. There are three main types of real-time PCR probes; TaqMan probes, Molecular Beacons and dual-oligo FRET probes. inqaba biotec synthesizes probes in all three classes. We have most of the 5’ fluorescent dyes as well as the 3’ quencher dyes. Following are tables of available fluorescent and quencher dyes.

*Dyes not available at inqaba biotec

The Black Hole Quencher (BHQ) dyes are a modern class of highly efficient dark quenchers that prevent fluorescence until a hybridisation event occurs. Adding a specific fluorescently-labelled hybridisation probe as the reporter instead of the DNA binding dye allows for the detection of only specific amplification products. The BHQ family readily permits single tube multiplexing due to the increased variety of reporter dyes that can be effectively quenched with little or no cross talk between reporters. Click here to download the BHQ and Dye selection Chart.

TaqMan PCR is at present, the most frequently used method for real-time PCR. The probes are longer than primers (20-30 bases long) with a Tm value of 10°C higher. They are designed to anneal to an internal region of a PCR product, and contain a fluorescent dye on the 5’ base and a quenching dye on the 3’ base. When irradiated the excited fluorescent dye transfers energy to the quenching dye (FRET). Close proximity of dye and quencher prevents emission while probe is intact. When Taq replicates a template on which the probe is bound, the 5’ exonuclease activity of Taq cleaves the oligonucleotide. Cleavage only occurs if the probe hybridizes to the target and the origin of the fluorescence is specific amplification. Specific requirement: No G must be present at the 5’ end as a G adjacent to the reporter dye quenches reporter fluorescence even after cleavage.

inqaba biotec Reporter – Quencher Pairs

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Molecular Beacons are hairpin-loop shaped single-stranded oligonucleotides consisting of a probe sequence that is homologous to the target sequence flanked by complementary ‘arm’ sequences which are homologous to one another and not the target sequence. A fluorescent reporter is covalently attached to one arm and a quencher to the other arm. In the absence of the target sequence the fluorescent reporter is held in close proximity to the quencher and any energy from the reporter is transferred to the quencher. When the probe bounds to its target, the greater stability of the probe-target helix forces the stem to unwind, resulting in a separation of the reporter from the quencher and now fluorescence can be monitored from the reporter molecule.

inqaba biotec Reporter – Quencher Pairs

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Dual-oligo FRET (Fluorescence Resonance Energy Transfer) Probes rely on the transfer of energy from one fluorescent molecule to another. Two separate sequence specific oligonucleotides are fluorescently labelled. The upstream probe has a donor molecule on the 3’ end and the downstream probe has an acceptor molecule on the 5’ end. Once the probes hybridize, the donor and acceptor fluorescent molecules are in close proximity to one another. This allows for the transfer of energy from the donor to the acceptor fluorophore, which emits a signal of a different wavelength. Either the decrease in the fluorescence of the donor or the increase in the fluorescence of the acceptor can then be detected. Thus, only when both probes are bound is the transfer of fluorescence detectable.

FRET probes allow for melt curve analysis and are useful in genotyping, SNP detection and other mutation detections.

inqaba biotec Reporter – Quencher Pairs

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DNA synthesis purification options

In DNA synthesis, each nucleotide is coupled sequentially to the growing chain. However in each coupling cycle, a small percentage (usually 1% or less) of the oligo chains will not be extended, resulting in a mixture of full length product and truncated sequences.

After the oligo is cleaved from the support and the protecting groups are removed, purification can separate the full-length product from the truncated sequence products. In general, the oligonucleotide purity required for a specific application depends on the potential problems from the presence of failure sequences (n-1, n-2…). For some applications, the presence of shorter oligos will not affect the experimental results. For other applications, it is imperative that only full length oligos be present. The table below lists the type of applications and the types of purifications recommended for these applications.

Some of the more common primer applications and the different purification types which we apply to different primer applications.

Type of application	Desalting	Cartridge	PAGE
Standard PCR (up to 30 bases)	X
Standard PCR (31 bases -35 bases)	_________	X
Standard PCR (36 bases -upwards)		_________	X
Real Time PCR incl. Molecular probes, Taqman probes, FRET probes			X
Degenerate primers		X
GC clamp primers	X
Primers for molecular cloning			X
DGGE primers	X		_________
FISH probes			X

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Delivery Time of Oligonucleotides

"¹Black Hole Quencher, BHQ-1, BHQ-2, CAL Fluor Orange 560, CAL Fluor Red 610, CAL Fluor Red 635, Quasar 570, and Quasar 670 are registered trademarks of Biosearch Technologies, Inc., Novato, California, U.S.A. (BTI) Patents are currently pending for the BHQ technology and such BHQ technology is licensed by the manufacturer pursuant to an agreement with BTI and these products are sold exclusively for research and development use only. They may not be used for human or veterinary in vitro or clinical diagnostic purposes and they may not be re-sold, distributed or re-packaged. For information on licensing programs to permit use for human or veterinary in vitro or clinical diagnostic purposes, please contact Biosearch at This e-mail address is being protected from spambots, you need JavaScript enabled to view it .

²TaqMan is a registered trademark of Roche Molecular Systems, Inc., Alameda, CA. PCR is a proprietary technology covered by several US patents including US Patent Nos. 4,683,195, 4,683,202 and 4,965,188, and by issued and pending counterparts outside the U.S. (PCR Patents). PCR Patents are owned by Roche Molecular Systems Inc. and rights under the PCR Patents have been obtained by Applied Biosystems in certain fields.

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