What is oligo scale?
What is oligo scale?
Oligonucleotides are synthetized in amounts (called scales hereafter) of 20 nmol, 40 nmol, 200 nmol, 1 µmol and 10 µmol. The ordered scale is an approximation of the amount of final product.
Do primers need to be purified?
Most site-directed mutagenesis protocols strongly recommend that you use only PAGE- or HPLC-purified primers to mutate plasmid templates. Using purified primers is supposed to minimize the introduction of unintended mutations, thus drastically improving the probability of generating your desired mutant.
How much do oligos cost?
|25 nmole DNA Plate Oligo||15 – 60 Bases||$0.19 USD / Base|
|100 nmole DNA Plate Oligo||10 – 90 Bases||$0.32 USD / Base|
|250 nmole DNA Plate Oligo||5 – 100 Bases||$0.54 USD / Base|
|1 umole DNA Plate Oligo||5 – 100 Bases||$1.06 USD / Base|
What does yield mean in PCR?
Yield, on the other hand, refers to the amount of final product recovered after all of the synthesis and purification steps associated with the oligonucleotide have been completed.
How do you purify an oligonucleotide?
Polyacrylamide gel electrophoresis is the oldest technique in oligonucleotide purification….The most common purification techniques used at Biosynthesis are:
- Polyacrylamide gel electrophoresis (PAGE)
- Reverse Phase High Pressure Liquid Chromatography (RP HPLC)
- Anion Exchange High Pressure Liquid Chromatography (AEX HPLC)
What is standard desalting?
Our standard desalting step refers to the removal of small organic molecules left over from synthesis. The desalting process does remove the majority of these organic molecules, though trace amounts can still be present in a shipped oligo.
How are oligos purified in the RPC process?
For oligos purified by RPC, the oligos are synthesized with the DMT group left on the final base, which allows for separation of impurities by the binding affinity of the hydrophobic DMT group to the resin in the cartridge, while truncated sequences missing their DMT are washed away.
How does the OPC oligonucleotide purification cartridge work?
The OPC® cartridge performs like an affinity column that adsorbs the dimethyltrityl (DMT) group of the trityl-on oligonucleotide while non-DMT bearing failure sequences, by-products, and other impurities wash through. The DMT group is removed with mild acid, allowing the purified, and detritylated oligonucleotide to be eluted.
Which is the best purification method for oligo primers?
Please refer to the purification options below for more information and suggestions. For unlabeled oligonucleotides (primers) Biosearch Technologies recommends RPC Purification which typically provides 70% purity*. Contaminants such as truncated sequences and other impurities are removed from the final product.
Which is the best purification method for oligonucleotides?
Reverse Phase Cartridge (RPC) purification is suitable to enrich the full-length product for oligos which contain 50 bases or less. For oligonucleotides containing modifications, such as fluorescent probes, we recommend more stringent purification such as single HPLC or dual HPLC.