What is DST purification?

During DNA manufacturing, each nucleotide is coupled sequentially to the growing chain via phosphoramidite chemistry. In each coupling cycle, a low percentage of the oligonucleotide chains do not extend, resulting in a mixture of full-length (n) and truncated (n-1, n-2, etc.)

What does an oligo do?

Oligonucleotides, or oligos, are short single strands of synthetic DNA or RNA that serve as the starting point for many molecular biology and synthetic biology applications! From genetic testing to forensic research and next-generation sequencing, an oligo may very well be the starting point.

What does HPLC purification do?

HPLC purification helps remove truncated synthesis products and enrich purity. IE-HPLC is useful for oligos that will be transfected into living cells, while RP-HPLC is needed to purify oligos with fluorescent dyes and some other hydrophobic modifications.

What are oligo primers?

The term oligonucleotide is derived from the Greek “oligo,” which means few or small. Oligonucleotides made up of 2′-deoxyribonucleotides are the molecules used in polymerase chain reaction (PCR). These are referred to as primers and are used to massively amplify a small amount of DNA.

Why do some pages purify primers?

Using purified primers is supposed to minimize the introduction of unintended mutations, thus drastically improving the probability of generating your desired mutant. However, specially purified primers can be extremely expensive, and take longer to synthesize than standard primers.

What are desalted primers?

Salt-free (“desalted”) oligos are suitable for applications such as microarrays, sequencing or qPCR primers. Reverse Phase Cartridge (RPC) purification is suitable to enrich the full-length product for oligos which contain 50 bases or less.

Are DNA oligos used for qPCR?

Oligo Designs Yes, you can perform real-time PCR / qPCR designs using our OligoArchitect Online tool.

Why are two oligonucleotide primers needed PCR?

Two primers are used in each PCR reaction, and they are designed so that they flank the target region (region that should be copied). That is, they are given sequences that will make them bind to opposite strands of the template DNA, just at the edges of the region to be copied.

What is dual HPLC?

Dual HPLC combines 2 HPLC purifications (RP+RP HPLC or RP+IEX HPLC) to obtain oligonucleotides with a higher purity level up to 95 %. This combined method allows to purif oligonucleotides up to 60 bases long, which could be used for stringent applications such as miRNA, siRNA, antisens and firststrand cDNA synthesis.

What are the two types of primer?

Types of Primers. There are three basic types of primers: oil-based, latex and pigmented shellac primer. Each has its strengths and weaknesses and works best on certain surfaces and in particular circumstances.

What is primer give example?

The definition of a primer is a coating that is applied to prepare a surface before the actual paint color is applied, or a textbook that is given to students to help them learn how to read. A sizing that is applied to a wall to prepare the surface for paint is an example of a primer.

How are oligonucleotides purified?

Hydrophobic oligonucleotides can be purified by ion-exchange HPLC by including an organic solvent in the mobile phase.

What are the impurities in the oligo purification process?

What Is the Oligo Purification Process? Oligonucleotide synthesis leads to the inevitable accumulation of impurities. These impurities are failure sequences: shorter sequences, longer sequences, modified sequences; or impurities left over by the synthetization process.

How does purification improve the performance of oligonucleotides?

For demanding applications such as multiplex PCR, cloning, mutagenesis or antisense/RNAi methods, additional purification can significantly improve oligonucleotide performance. Purified oligos up to 60 bases that are provided with a purity guarantee receive QC by capillary electrophoresis (CE).

How is reversed phase HPLC used in the purification of oligos?

Reversed-phase HPLC is used routinely in the purification of oligos. Oligos with a high degree of secondary structure (e.g. hairpin loops) give rise to multiple peaks. This secondary structure can be eliminated in the presence of a highly alkaline pH, which eliminates hydrogen-bonding interactions.

When do you need a dual labeled oligo?

Sometimes a labeled or dual labeled oligo is required for molecular beacon, FRET, biotin-streptavidin, or bio conjugation applications. In these special cases, the hydrophobicity of the modifying group at either the 5’ or 3’ end can be utilized to produce high quality oligos through C18 reverse phase HPLC.