LightCycler®/HybProbes - LCPs
Following the concept of fluorescence resonance energy transfer (FRET), metabion synthesizes LightCycler® (LC®) probes in an ISO-certified manufacturing facility using processes licensed by Roche. We offer LC®probes in 7 different scales at defined molar amounts.
Introduced to the market in 1997, Light Cycler technology has become increasingly popular in various areas, from research to diagnostics.
The LightCycler® hybridization probe system is based on two oligonucleotide probes that hybridize, in a head-to-tail arrangement, to adjacent sequences of target DNA (hybridization assays). Each probe carries a different marker dye, the first 3'Fluo, the second 5'LC®610, 5'LC®640, 5'LC®670 or 5'LC®705. In addition, the 3'OH terminus of the LC® probes is "capped" by a phosphate group to prevent the polymerase from extending the primer during the PCR reaction.
As the marker dyes are in close proximity, fluorescence resonance energy transfer between them takes place which allows a specific and quantitative real-time detection of the amplification of a nucleic acid template. To avoid sterical problems between the 3' Fluo and 5' LC® Red end labels of the probes there should be a spacer of 1 to 5 nucleotides (4 to 25 Å distance) to separate the two probes from each other.
metabion synthesizes LC® probes in an ISO-certified manufacturing facility using processes licensed by Roche. We offer LC®probes in 7 different scales at defined molar amounts.
Our LightCycler/HybProbes - Portfolio
- These fluorophores can be positioned internally and at the 5' or 3'-end, to produce dual or multiple labelled probes. To learn more click here.
- We offer LC® probes with a maximum length of 40 nucleotides in guaranteed delivered quantities between 1 and 30 nmol. Our service includes HPLC and Mass Check. Click here!
- For larger quantities, contact us!
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 LC®probe/s.
- Sequence of the LC®probe/s in 5’-3’ orientation.
- Yield range.
- Purification required.
- 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).
The expected average in-house turnover time is 4–5 working days. Please note that we perform strict quality controls on your ordered LC®probe. In case one or more LC®probe do not pass our quality control, they will have to be resynthesized. This may, of course, result in a delay.
The label on the probe tube shows basic information like oligo name, name of person who ordered, probe sequence including modifications, probe ID, amount of DNA (OD260 and nmol), Tm, and molecular weight.
In addition, you will receive a synthesis report 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. LC®probes are always HPLC purified and you will also get a printout of the preparative chromatogram.
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 and you will receive a hard copy of the Mass Check spectrum.
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.
For a detail explanation of LC® probe and their use in qPCR, please refer to our Knowledge-hub.
You also need to consider the following:
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 LC®probe may vary from powdery to hyaloid. The color changes according to the dye attached.
Purified distilled 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 (see also FAQ "How stable is my probe once I have resuspended it?").
Standard concentration for PCR primers is 0.1 mM.
To gain a maximum shelf life for oligonucleotides, samples should generally be stored dehydrated at ≤ –15 °C in absence of light. Under the mentioned conditions, samples are stable for at least 6 months. In case of a longer storage period, oligos should be pretested for molecular integrity prior to experimental use. If a sterile solution (e.g. water, biological buffer) is used as diluent, the re-suspended the probe will be stable at 20 °C for several days to weeks, at 4 °C for about a month. If stored frozen at –20 °C or –70 °C, it will remain stable for several months.
For correct storing and best performance of your probe, we recommend the following:
- Avoid repeated freeze-thaw, as this will denature the probe;
- Avoid the use of distilled water as a diluent, since its pH may be as low as 4–5. The probe stability in solution depends on the pH. Dissolving probes into acidic solutions may result in oligo degradation. Therefore, use purified distilled water;
- Minimize the exposure of fluorescent probes to light, to avoid any bleaching effect;
- Store probes 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.