RNA qPCR probes
metabion's RNA dual labelled probes (RNA DLPs) are optimized for use in real-time quantitative PCR and produced according to technologically advanced synthesis and purification protocols to reduce background noise to an absolute minimum.
Our range of reporter-quencher combinations suits both probe hydrolysis- and hybridization-based assays. We believe this will cover your needs for single or multiplex real-time PCR applications.
We offer our RNA DLPs in final yields in nmoles. With this, we introduce a transparent and best price-performance scheme for our broad selection of dual labelled probes as follows:
- 5 scales based on delivered quantities tailored to your routine needs;
- length-independent guaranteed (range) of delivered quantities in nmoles for probes from 15 to 40mers. Please note that OD260 values are a measure of total nucleotides´ optical density. Hence, neither purity nor amount of ordered substance is transparently reflected. For simplification and exemplification purposes, consider the following:
1 OD of the 20mer 5´CAT CGT ATT CGA TGC TAC GT 3´ translates into approximately 5 nmol;
1 OD of the 40mer 5´CAT CGT ATT CGA TGC TAC GT CAT CGT ATT CGA TGC TAC GT 3´ translates into approximately 2.5 nmol.
Therefore, a 1 OD guaranteed amount of delivered product can vary significantly, while metabion's commitment to delivered yields in nmol for probes ranging from 15 to 40 nucleotides does not allow for ambiguity in terms of what you expect and pay for.
- metabion's routine dual-labelled probe portfolio perfectly covers the spectrum of applicable excitation and detection wavelengths, with reporter-quencher combinations that are suitable for all commonly used real-time PCR platforms. However, do not hesitate to ask for non-listed combinations including internally or multi-labelled oligos and probes. Contact us.
- Our Prices include HPLC purification as well as Mass-Check QC for our high-quality products.
- You will receive your RNA DLPs under dry shipping conditions, unless otherwise specified.
It matters what you actually get for your money!
Our standard custom RNA qPCR probes - Portfolio
5 yield ranges based on independent lengths of delivered oligonucleotide quantities from 1 to 40 base length:
- ≥ 1 < 5 nmol
- ≥ 5 < 10 nmol
- ≥ 10 < 20 nmol
- ≥ 20 < 30 nmol
- ≥ 30 < 50 nmol
All our RNA DLP probes are HPLC purified and mass-checked for best quality assurance! Please note that these services are included.
FAQ – you ask, we answer
There are two ways of ordering:
- The preferred way is order transmission through our Web Order Portal for most convenient online shopping (please refer here for further details).
- You can order by sending us an e-mail at email@example.com with our pre-formatted excel order file as attachment. Download respective Order Form.
When you write your e-mail, please make sure to include the following required information in the excel template:
- Name of the RNA DLP/s.
- Sequence of the RNA DLP/s in 5’-3’ orientation.
- Yield range.
- Delivery form (dry/in water/buffer + concentration).
If you are a new customer, please additionally provide us with:
- Your shipping and billing address.
- Any other information like Purchase Order number, VAT number (VAT only for customers resident in the EU), etc.
In case you choose to transmit orders via e-mail using your own format(s), we need to alert you that above mentioned information in bold print is obligatory for processing your order. Due to extra efforts necessary for individual order format transfer into our system, order processing will take longer as compared to preferred web orders and pre-formatted e-mails. Please note that only files updated to the latest excel/word version (e.g. .xlsx or .docx) are accepted.
How can you convert files to newer formats?
Open the file in a recent Office program and save the file in a newer format. For this go to “File” → “Save as” and choose the newer format from the “Save as type” dropdown menu.
If you want to connect your eProcurement System with our Web Order Portal (e.g. OCI - Open Catalog Interface), please simply contact our Customer Service (firstname.lastname@example.org).
Using our optimized production pipeline, we can deliver RNA DLPs of over 40 bases. The maximum length of an oligonucleotide depends both on the sequence and on the modifications of interest. For example, consider a DLP having a reporter at the 5’-end and a quencher at the 3’-end. The length of the DLP can compromise the activity of the quencher, if the distance between reporter and quencher is too high. Therefore, we consider a length of 40 bases as maximum, for a DLP probe to function optimally.
To get best performance for your experiments, we can help you designing your probe. Please do not hesitate to contact us!
5 yield ranges based on length independent delivered quantities* for probes from 18-40 nucleotides:
- ≥ 1 < 5 nmol
- ≥ 5 < 10 nmol
- ≥ 10 < 20 nmol
- ≥ 20 < 30 nmol
- ≥ 30 < 50 nmol
- ≥ 50 nmol
* Please note that the term “yield range” refers to the final amount of product you will actually receive. Instead, OD260 values are a measure of total nucleotides' optical density. Hence, neither purity nor amount of ordered substance are transparently reflected. For simplification and exemplification reasons look at the following:
1 OD of the 20mer 5´cau cgu auu cga ugc uac gu 3´ translates into approximately 5 nmol;
1 OD of the 40mer 5´cau cgu auu cga ugc uac guc auc gua uuc gau gcu acg u 3´ translates into approximately 2.5 nmol.
For a detail explanation of DLPs and their use in qPCR, please refer to our Knowledge Hub.
You also need to consider the following:
The expected average in-house turnover time is 10–15 working days. Please note that we perform strict quality controls on your ordered RNA DLPs. In case one or more RNA DLPs do not pass our quality control, they will have to be resynthesized. This may, of course, result in a delay.
The label on the RNA DLP tube shows basic information like oligo name, name of person who ordered, DLP sequence including modifications, RNA DLP oligo ID, amount of DNA (OD260 and nmol), Tm, and molecular weight.
In addition, you will receive a technical data sheet 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. Additionally, you will receive a hard copy of the Mass Check spectrum.
The following terminology is used for differentiating between offered QC options including respective documentation coverage in our order forms and on supporting documents delivered with the products:
Standard quality control performed on each and every oligo. Either MALDI- or ESI-ToF, subject to the "nature of the oligo", and metabion internal procedures. This service is free of charge.
Mass Check + Analytical HPLC
Explicitly ordered and performed Mass Check (MALDI-ToF or ESI-ToF subject to the "nature of the oligo" and metabion internal procedures) and Analytical HPLC (see FAQ: "What is the difference between preparative and analytical HPLC?"). Product delivered with analytical HPLC and MS spectra. Additional charges apply.
Maintaining sterile, RNAse free conditions is always recommended as a precaution. Dried pellets are stable at room temperature for 2–4 weeks, but should be placed at –20 °C or –80 °C for long term storage. Under these conditions, dry RNA is stable for at least one year.
For correct storing and best performance of your RNA DLP, we recommend the following:
- Avoid repeated freeze-thaw, as this will denature the RNA DLP;
- Avoid the use of water as a diluent, since its pH may be as low as 4–5;
The RNA DLP stability in solution depends on the pH. Dissolving DLPs into acidic solutions may result in oligo degradation. Therefore, avoid the use of non-sterile distilled water as solution pH might be as low as 4–5;
- Minimize the exposure of RNA DLPs to light, to avoid any bleaching effect;
- Store RNA DLPs highly concentrated and not in working dilutions, if you are not planning to use them within 24 hours. The higher the dilution factor, the faster the fluorescent activity fades away. Therefore, try to store highly concentrated aliquots frozen, thaw them only once, dilute them just before you use the probe and store the aliquots at 4 °C in the dark;
- For general instructions on how to handle RNA, please refer to our Guidelines for handling RNA.
Distilled sterile water pretreated with DEPC, or any nuclease free biological buffer (i.e. with physiological pH) are acceptable as diluents. The recommended diluent volume is 100 µl - 1 ml, the concentration depending on the application to be used and the yield of the resulting product (see also FAQ "How stable is my RNA DLP once I have resuspended it?").
Please see also our RNA-handling guidelines!
There is a normal degree of variation in the appearance of the supplied dry oligonucleotide pellets. Variation in appearance per se does not indicate a quality defect. In general, appearance of DLPs may vary from powdery to hyaloid. The color changes according to the dye attached.