- What chemistry is used for synthesis of metabion's RNA Oligonucleotides?
- What purification methods are used for RNA oligonucleotides?
- What purity level can I expect?
- Is there a maximum length for synthesis of RNA oligonucleotides?
- What is the coupling efficiency for RNA oligonucleotides?
- Which kind of quality control is used for the RNA oligonucleotides?
- What synthesis scales are offered for RNA oligonucleotides and how much RNA will I get?
- In which form do I receive my RNA oligonucleotides?
- How can I tell how much RNA I have received?
- Which modifications can be used for RNA?
- Can I get an RNA oligo with a dideoxynucleotide at the 3' end?
- Do you offer other RNA-DNA chimera?
- How should RNA oligonucleotides be stored?
- How stable is the RNA?
- What should I dissolve my RNA pellet in, water or buffer?
1. What chemistry is used for synthesis of metabion's RNA Oligonucleotides?
The chemistry of RNA synthesis is identical to the DNA synthesis except for the presence of an additional protecting group at the 2' hydroxyl position of ribose. This position is protected with silyl groups (usually tBDMS), which are stable throughout the synthesis. The remaining positions on both the sugar and the bases are protected in the same fashion as in DNA.
2. What purification methods are used for RNA oligonucleotides?
Our default purity is HPLC! Any RNA oligonucleotide shipped to you is purified for full-length to guarantee the top quality required for your experiments. HPLC purification is included in our price per base.
3. What purity level can I expect?
≥ 95%!
4. Is there a maximum length for synthesis of RNA oligonucleotides?
The current maximum length of RNA oligonucleotides is 80 bases.
5. What is the coupling efficiency for RNA oligonucleotides?
98-99%
6. Which kind of quality control is used for the RNA oligonucleotides?
Synthesis is monitored through trityl analysis. Quantitative HPLC purification and final spot checks by mass spectroscopy further ensure top quality.
7. What synthesis scales are offered for RNA oligonucleotides and how much RNA will I get?
| Scale (µmol) | Unmodified oligos Yield* (OD260) HPLC Purified |
5' or 3'modified oligos Yield* (OD260) HPLC Purified |
| 0. 2 | 8 | 3-5 |
| 1.0 | 18 | 15 |
*Guaranteed yields apply for a 20mer +/- 20%! For oligos >33nt we cannot give a yield guarantee!
8. In which form do I receive my RNA oligonucleotides?
Lyophilized.
9. How can I tell how much RNA I have received?
A comprehensive Synthesis Report comes along with each order, indicating oligo name and sequence, composition of bases, modification, synthesis scale, yield in ODs, µg, nmol, Tm and molecular weight.
10. Which modifications can be used for RNA?
5' modifications: HEX, FAM, TET, TAMRA, Fluorescein, Biotin, Phosphate, Amino, CY3, CY5
3' modifications: TAMRA, Biotin, Amino, Thiol
11. Can I get an RNA oligo with a dideoxynucleotide at the 3' end?
Yes.
12. Do you offer other RNA-DNA chimera?
Yes.
13. How should RNA oligonucleotides be stored?
The best way to store RNA is as a dry pellet at -20°C or colder. Avoid frequent freeze-thaw cycles
14. How stable is the RNA?
As a dry pellet the can be stored at -20°C for 6 months.
15. What should I dissolve my RNA pellet in, water or buffer?
We recomend to dissolve the single stranded RNA in 1X TE buffer (prepared under RNase-free conditions (10 mM TrisCl, pH7.5, 0.1 mM EDTA)). This buffers the pH and chelates metal ions that can contribute to RNA degradation. RNase-free water is also acceptable.
metabion's RNA oligonucleotides are delivered deprotected and purified. They are RNAse free, but as RNA is highly susceptible to degradation by exogenous RNAses introduced during handling, it is essential that you conduct all handling steps under sterile, RNAse free conditions. Never handle RNA without wearing gloves. RNAse free reagents, barrier pipette tips and tubes should be used!