RNA Oligos - plates

There are two ways of ordering:

1. The preferred way is order transmission through our Web Order Portal for most convenient online shopping.
2. You can order by sending us an e-mail with our pre-formatted excel order file as attachment. Download respective Order Form
   

   RNA Oligos - plates	download xlsx
   Also available in our web order system (WOP)

   When you write your email, please make sure to address the following questions in the excel template:
   
   
   • Name of the oligos?
   • Plate type?
   • Position of the oligo?
   • Range of delivered quantity
   • Modifications?
   • Sequence of the oligo in 5’-3’ orientation?

   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

Jump on our Web Order Portal (WOP), look for the new product category headline under “Custom Synthesis Services” called “High Throughput Oligos (HTOs)”, and open the respective order form and start filling it. Options provided are self-explanatory. The system shall guide you through the ordering process.

Place your order and benefit from our newly introduced yield scale and pricing scheme.

Confirmation

Online orders or email orders which indicate an email address will be confirmed by email.

Delivery

Our default shipping mode is sending by Express service overnight at EURO 4.20 per shipment within Germany. If the value of your order is > EURO 125,00 shipping within Germany is free of charge!
For other countries please see our shipping table.

Average in-house turnover times (freight forwarders delivery time not included):

• RNA oligos in plates: 5-7 business days

Above mentioned estimated turnover times are only indications and refer to our standard portfolio. In terms of "counting" working days, orders placed past 15:00 (Munich time) are considered to be next day's order. Major deviations will be communicated timely. Be assured that we try to process your order as quick as possible without compromising on quality!

All our products are offered according to our terms and conditions.

2 ranges of quantity delivered based on length independent delivered quantities* from 10-50 nmol for oligos from 5-40 nucleotides:

• ≥ 10 < 25 nmol
• ≥ 25 < 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 DNA 20mer 5´cau cgu auu cga ugc uac gu 3´ translates into approximately 5 nmol.

1 OD of the DNA 40mer 5´cau cgu auu cga ugc uac guc auc gua uuc gau gcu acg u 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 oligonucleotides in nmol does not allow for ambiguity in terms of what you expect and pay for, not even for modified oligos!

For more information, click here.

Standard modification pricing refers to one modification per oligonucleotide. Dual or multiple labelling does not automatically follow simple addition arithmetics. Please inquire for feasibility and pricing.

Synthesis scale refers to the amount of starting CPG (controlled-pore glass) support-bound monomer used to initiate the DNA synthesis, not the amount of final material synthesized. This is the same for all manufacturers of synthetic DNA using standard phosphoramidite chemistry. When a synthesis scale of 40 nmole is specified, approximately 40 nmoles of the first base are added to the DNA synthesizer. For an average 25-mer, at least 25% of this starting material will result in failure sequences; hence it is not possible to produce 40 nmoles of full-length product from a 40 nmole scale synthesis. The losses occur during synthesis, post-synthetic processing, transfer of material, and quality control. Final yield is the actual amount that we guarantee to deliver.

Please note that 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´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 does not allow for ambiguity in terms of what you expect and pay for.

Expected turnover time for

• Desalted RNA oligonucleotide in single tube format is 3-5 days.
• RNA in plates, expected turnover time is 5-7 business days*.
• HPLC-purified, unmodified or single-labelled RNA oligonucleotides from our standard portfolio are usually processed within 5-10 business days including purification and quality control*.
• Duplex oligos take about 10-15 days to be produced, purified and QCed*.
• For RNA dlps you should consider 10-15 days as a realistic turnaround time and for RNA longmers depending on sequence length/complexity and modification level 8-15 business days*.

*Freight forwarders delivery time to the final destination is not included in all cases. In terms of "counting" working days, orders placed past 15:00 (Munich time) are considered to be next day's order. Be assured that we try to process your order as quickly as possible without compromising on quality!

Of course we can and we would be happy to do so! Please inquire for feasibility and a quotation; write us an email to info@metabion.com.

This depends on the complexity (length, base composition, modifications) of your requested molecule, as well as on the application desired. Failure sequences may be generated both during and post-synthesis. Due to the nature of synthesis chemistry (coupling efficiency < 100%) and/or post-synthetic modification procedures, there will remain failure sequences (n-x), free modifiers and non-labeled product in the "crude" unpurified product.

We strongly recommend that all modified oligos and oligos longer than 80 nucleotides are subjected to HPLC purification, no matter which kind of downstream application is envisaged.

For "critical" applications like gene synthesis, subcloning, mutagenesis and alike, HPLC purification should be considered for oligos > 45 nucleotides.

HPLC purification is standard (no additional charge) for all our dual-labeled probes and/or multi-labeled oligos, as well as for all our RNA and large scale oligos.

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.

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 unmodified and dye-labeled oligo pellets may vary from powdery to hyaloid. The color of unmodified oligo pellets may range from transparent over off-white and yellowish to tan. The pellets of labeled oligos are colored according to the dye attached.

Purified water, TE or any biological buffers (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. Standard concentration for PCR primers is 0.1 mM.

Metabion is dedicated to reliably deliver high quality products. While every production step is performed in light of achieving best quality, the product is released only if it passes our final inspection. Mass Spectrometry has become the state-of-the-art technology for verifying the integrity of oligonucleotides, and metabion has been the first custom oligo house who introduced routine mass checks into its operations. Each and every oligo is characterized by either MALDI- or ESI-ToF and stringent release criteria are applied.

Mass Spectrometry allows for the most sensitive detection of low-level by-products/impurities such as

• n-1/n-x oligos
• Depurination
• Incomplete Deprotection
• Acrylonitrile adducts
• High Salt Content Identification

Moreover, it is the fastest and most efficient way to identify potential product mix-ups.

We run two different types of Mass Spectrometry (MS) instruments in order to cope best with quality and quantity/throughput issues determined by the specifications of the respective oligo/analyte. While each instrument type precisely characterizes oligonucleotides in terms of composition through direct molecular weight measurement, their field of application is diligently adjusted to suitability considerations.

MALDI-ToF instruments typically have a higher throughput, while the limits of using this technique become manifest, if it comes to analyzing long oligonucleotides, or oligos carrying certain photo-labile modifications (e.g.common quenchers like BHQ®s, Dabcyl used in DLPs).

ESI-ToF is less efficient in terms of throughput but perfectly compensates for resolution issues with long oligos as well as for a potential detrimental laser impact on labile/photosensitive modifications – thus being a "natural" complement to MALDI-ToF analysis.

Synthetic oligonucleotide purification is particularly challenging because of the small differences in size, charge and hydrophobicity between the full-length product and impurities, which often co-elute.

For improved analysis of complex samples like long and/or multiple labeled oligos, metabion offers liquid chromatography (LC) coupled with electrospray ionization mass spectrometry (ESI-MS). The mass spectrometer is connected to a high pressure liquid chromatography (HPLC) system, which allows premium analyte characterization via chromatographical separation, followed by respective molecular weight determination. With this system, the mass of oligonucleotides between 2 and 220 bases can be analysed with high accuracy, resolution and sensitivity. Our expert production team will take care of the method (MALDI or ESI ToF) that best applies to your sample.

Preparative High Pressure Liquid Chromatography (HPLC) deals with isolating the separated components of a sample, and can be done on small-, mid- and large scale operations. In other words, the objective of a preparative HPLC is isolating and purifying a product. Practically, the sample goes from the detector into a fraction collector or it is collected manually.

Analytical HPLC refers to the processes of separating and identifying the components of a sample. It is usually a small-scale process, whose objective is the qualitative and quantitative determination of a compound. The sample goes from the detector into waste.

metabion offers analytical HPLC as an additional (optional) quality control method, complementing our Mass-Check QC, which is performed by default on all our oligos.

For product/quality documentation please see FAQ: What kind of documentation do I get with my RNA oligos?

The label on the oligo plate shows basic information like name of person who ordered, name of the plate and order ID.

In addition, you will receive a synthesis report containing more detailed information, such as name of the oligos, physical-chemical properties of the oligos, such base composition, base count, purification grade, amount of DNA (OD₂₆₀ and nmol), Tm and molecular weight.

While each and every oligo produced and delivered is characterized by either MALDI- or ESI-ToF before release, Mass-Check documentation/traces will only be provided if requested at the time of order placement. Additional charges may apply. 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:

Mass Check
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 and no printed/pdf documentation is provided.

MALDI-ToF
Explicitly ordered and performed MALDI-ToF check. Product delivered with MALDI-ToF traces. Additional charges apply.

ESI-ToF
Explicitly ordered and performed ESI-ToF check. Product delivered with ESI-ToF traces. Additional charges apply.

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´s the difference between preparative and analytical HPLC?). Product delivered with analytical HPLC and MS traces. Additional charges apply.

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. Please visit our DNA FAQs for more information.

For RNA oligonucleotide between 5-40 nts and for a quantity delivered between 10-100 nmol, we offer RNA oligonucleotides desalted as well as HPLC purified. Over 100 nmol and 40 nts our default purity is HPLC! Such RNA require HPLC purification to guarantee the top quality required for your experiments. For such RNA oligonucleotides, HPLC purification is included in our price per base/probe/duplex. Visit our Standard Portfolio.

Synthesis is monitored through trityl analysis. Quality check by mass spectrometry further ensure top quality. Please visit our DNA FAQs page for more information.

Our default delivery form is dry. This ensures maximum stability during transport and storage.

The best way to store RNA is as a dry pellet at -20°C to -80°C and to avoid frequent freeze-thaw cycles. If you want to store your RNA in solution re-suspend the delivered pellet in an RNAse-free solution buffered at pH 7.4 - 7.6 and store at -20°C or less. We recommend that RNAs are re-suspended at a convenient stock concentration and stored in small aliquots to avoid multiple freeze thaw cycles.

For modified oligonucleotides – especially for fluorescent dye labelled oligos – you should minimize the probe’s exposure to light because of its bleaching effect. Additionally, we recommend storing dye labelled oligos highly concentrated, rather than in working dilutions, if you don’t use them immediately. The higher the dilution factor the faster the fluorescent activity decreases/fluorescence fades away. Therefore, try to store highly concentrated aliquots frozen, thaw them only once, and dilute them just before usage. Click here to download our guidelines for handling RNA.

We recommend to dissolve single stranded RNAs in 1x TE buffer (10 mM TrisCl, pH7.5, 0.1 mM EDTA; prepared under RNase-free conditions). This buffers the pH and chelates metal ions, which may contribute to RNA degradation. RNase-free water is also acceptable.

metabion's RNA oligonucleotides are delivered deprotected and purified – ready to use. 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!

For more information, please read FAQ: How should siRNA oligonucleotides be stored?

You should also read our guidelines.

RNA duplexes are significantly more resistant to nucleases than a single strand RNA oligonucleotide. However, 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.

A comprehensive Synthesis Report comes along with each order, indicating oligo name and sequence, composition of bases, modification, base count, purification, yield in ODs, nmol, Tm and molecular weight.

For desalted RNAs falling into the specifications of our standard portfolio, you can expect at least >80% of purity. HPLC-purified RNAs falling within the specifications of our standard portfolio, have typical purity >85%. Please note that most delivered RNAs will have purities >90%.

All oligonucleotides, whether single-stranded or double-stranded oligos, are provided as dried pellets and shipped at ambient temperature. While being stable at room temperature for 2-4 weeks, they should be placed at -20°C or at lower temperature upon receipt.