This may be useful for sequences withlow primer melting temperatures.

Unless requested, oligos are synthesized without either 3´or 5´ phosphate. The 5´ and/or 3’-phosphate is available as a modification at additional charge.

Use the PCR buffer with a workingmagnesium concentration of 1.5 m and 2m primers.

The label on the oligo tube shows basic information like oligo name, name of person who ordered, oligo sequence including modifications, oligo ID, amount of DNA (OD260 and nmol), Tm, and molecular weight.
In addition, you will receive a synthesis report containing more detailed information on the physical-chemical properties of the oligo, such as base composition, base count, purification grade, amount of DNA (OD260 and nmol), Tm and molecular weight. If you have ordered purification or your ordered oligo is purified by default (e.g. Dual labeled oligos, dual modified oligos, mid- and large scale oligos), you will also get a printout of the preparative chromatogram.


Designing PCR primers and probes

Typically by startingwith 2 n of synthetic pool oligonucleotide and ~2 m

Chemically synthesized oligonucleotides may contain a number of modificationsincluding deletions, insertions, incompletely deprotected bases, or backbonelesions all of which affect PCR efficiency.


Biolegio | Custom Oligos Synthesis

As with plasmid DNA templates, PCR products should be of sufficient quality toensure successful sequencing. It is essential to remove potentially interfering substancesthat may remain in solution after the PCR reaction. Potential contaminants include excess PCRprimers, nucleotides, enzyme and buffer components from the PCR reaction as well as unspecificamplification products. These contaminants can sometimes participate, and thus interfere,in the cycle sequencing reaction and lead to poor quality data or no data at all.

Please consider the following recommendations when preparing your PCR reaction(s) for submission:

Large Scale Oligo Synthesis; Optional Services

metabion offers standard synthesis of Mid/Large scale DNA oligos between 15 and 25 bases of length. Our customers can select a final yield between 50 mg and 1 gram. For modified sequences and lengths outside of the standard range above stated, please send your inquiry to

PCR and qPCR Primer Design Tutorials with PrimerQuest

For example, if the 1/64 dilution is the first to showno detectable DNA (implying that 6 doublings of the synthetic DNA tookplace yielding 64 fold more DNA) and if 7 cycles of PCR were performed,then the average number of doublings per cycle is ~1.81.

as a primer to initiate template- directed DNA synthesis in PCR ..

For example, if the 1/64 dilution is the first to showno detectable DNA (implying that 6 doublings of the synthetic DNA tookplace yielding 64 fold more DNA) and if 7 cycles of PCR were performed,then the average number of doublings per cycle is ~1.81.