Primer synthesis

A brief introduction of primer synthesis

At present, primer is always synthesised by using solid phase and phosphoramide three ester method. Using this method to synthesis DNA fragments with efficient, fast coupling and the characteristics of the initial reactant stable. And the method is fixed DNA strands on solid phase carrier to proceed DNA synthesis, primers was synthesized in the direction of the 3 'end to the 5' end, the adjacent nucleotides connect by 3 '- 5' dihydrogen phosphate ester bond.

Synthetic process

The first step is to react protected nucleotides connect in the active group on the solid phase carrier in advance with trichloroacetic acid, take off the DMT 5 '- the protection of the hydroxyl groups, get the 5' - hydroxyl free. The second step, synthesis raw material for the DNA, and phosphoramidite protect the nucleotide monomers, mixed with activator four azole nitrogen, get nucleoside phosphite activation intermediates, it’s the 3 'end actived, 5' - is still DMT hydroxyl protection, with the 5 '- free hydroxyl in the solution condensation reaction. The third step, with capping reaction, condensation reaction may have a small number of 5 '- hydroxy (less than 2%), did not participate in the reaction, with acetic anhydride and 1 - methyl imidazole terminate continue to react, this short fragments can choose separation purification way off according to the experiment in the subsequent. The fourth step, under the action of oxidant iodine, the phosphorus acyl formed into a more stable phosphate ester.

Through the four steps above , a DNA nucleotide is attached to the solid phase carrier. By trichloroacetic acid off its 5 '- hydroxyl protection group on the DMT, repeat the above steps, synthetic base pick up until all the requirements. Can observe the color of TCA processing stages in the process of synthesis judge synthetic efficiency. Through ammonia water high temperature treatment, primer connected on the CPG was cut off, purified by OPC, PAGE methods, product primers with C18 enrichment, desalination, precipitation. After precipitation primers was suspended with water , determine OD260 quantitative, partial shipments according to the order requirement.

Service features and advantages

Strict quality control, high purity quality guarantee: Trigoats have a variety of synthetic instrument, which is dominated by BLP platform and SpectraMax collocation, used in the synthesis of high flux, monitored synthesis efficiency. Also we own ABI 3900 and BioAutomation used in large-scale synthesis, specialized for the synthesis of some special purpose. In addition, the company has a variety of models of high performance liquid chromatograph, can meet the level of 50 nmol - 50 umol synthesis, purity can reach more than 95%.

The delivery is fast

Common primers within 48 hours of delivery, the domestic 1-2 working days. DSL and OPC purification primers (below 60 base) delivery time 1 to 2 working days commonly, PAGE and HPLC purification primer delivery time general 2-3 days, modified primer 5-7 delivery is commonly 3-5 working days. Company with gas phase ammonia opv-la aminolysis with high temperature and high pressure, avoiding the ordinary ammonia solution, speed up the ammonia solution, reduce the trace substances residues and pollution in the process of the ammonia solution, the damage of primers down to the degree. Besides, we adopt the automatic control of ammonia solution, reduce the accidental deviation in the process of ammonia solution.

A variety of purification methods

DSL purification/C18 desalination and purification; OPC purification; PAGE purification; HPLC purification; Using the freeze-drying technology, high stability of products. We adopt freeze-drying technology to sythesis the primers of high amount of OD, compared to peer vacuum heating dry, can greatly reduce the damage to the product. Freeze-dried primers in - 20 ° C can be stable for a year, in 4 degrees can be stable for several months.

Synthesis of a variety of modified primer

Our company can provide a variety of high quality fluorescent decoration, including double marking fluorescent probes, rare bases, phosphorothioated Oligos, phosphate groups, all kinds of fluorescent group, biotin, digoxin, etc.

Powerful technical support

Solve any problems of the primer at any time for you.