Hello, I would like to ask you if it is possible to use CleanTag Small RNA library prep kit also for piRNA sequencing or if the adapters bind only to miRNAs? -Best, Petra

Hello Petra,

CleanTag library preparation is a ligation based method that is designed to tag adapters onto small RNA species with a 3′ OH and 5′ P. piRNA is thought to have this type of structure and would therefore be compatible with the CleanTag kit.

Best Regards,

Joey

What are the minimum requirements for shipping mRNA over a 24-48h period? Is there a minimum volume that it should be shipped at e.g. not less than 100 ul? Could a lower volume mean the mRNA is more susceptible to degradation? Are there certain buffers that you always recommend? Is there an easy assay that could be carried out each time the mRNA is used to determine whether any loss in quality has occurred? Many thanks-Heather

Dear Heather,

Thank you for the inquiry. We routinely prepare aliquots of our mRNA in sizes as small as 20 uL @ 1 mg/mL, although we do not recommend aliquot sizes any smaller than 20 uL. We also ship all of our mRNA products in an insulated box on dry ice, and recommend storage at -40°C or below.

We formulate all of our catalog mRNAs in 1 mM Sodium Citrate, pH 6.4. We have found that this buffer provides both high solubility and stability of the mRNA when stored and handled properly. If you have identified a different buffer that you prefer, we can likely formulate the mRNA in that buffer for you as an alternative. We do recommend that single-use aliquots be prepared in order to minimize freeze-thaw cycles, which can cause degradation. To analyze the mRNA prior to use, we recommend confirming the length of the mRNA on a gel and/or bioanalyzer.

Best Regards,

Scott

We need to quickly move forward with our novel therapeutic oligo. Can TriLink produce clinical grade material and quantities?

Yes! For over 20 years, we’ve been industry leaders synthesizing high quality research and diagnostic grade material, especially difficult and unusual constructs. We combine our expertise with a state-of-the art GMP manufacturing facility to offer mRNA, oligonucleotide and small molecule therapeutic development and production services, tailored for your specific applications.

TriLink can ensure methods and materials are compatible with GMP manufacturing requirements from the start of your research project. From idea conception to early human trials, TriLink is an expert partner. We offer consultation on compound design, material grade, process development, scale up and cGMP production. TriLink delivers consistent quality API on schedule, regardless of the scale. Plus, TriLink provides supporting documentation to streamline the IND submission process and carry you through to clinical trials – making the process efficient and cost effective.

TriLink has successfully taken targets from small, research grade syntheses through to large scale cGMP batches, including upwards of 10 grams of mRNA therapeutic compounds. We offer a wide range of manufacturing scales enabling you to specify the exact amount of material required for IND-enabling tox studies and early phase clinical studies.

With nine dedicated GMP manufacturing suites and two support labs, TriLink has the agility to meet most timelines by implementing and managing multiple projects in parallel. Our PMP certified project management team will ensure your product is delivered on time and on budget. Open communication and regularly scheduled updates throughout the manufacturing process will keep you  apprised of progress.

We know drug development is complicated, leverage our experience to ensure your success.

Request a GMP Consultation today:

What is the difference between your ChromaLink™ Kit and your ChromaLink™ One-Shot Kit?

The ChromaLink™ One-Shot Biotin Kit (B-9007) biotinylates 100 µg of antibody at 1 mg/mL. We also have an additional kit, the ChromaLink™ Biotin Labeling Kit (B-9007) which is more flexible in format and can label up to 5 reactions of between 25 µg to 1 mg per 100 µL reaction. The reaction conditions were specifically designed to match the concentrations and quantities of most commercially available antibodies.

Dear sir/madam : I am planing to do selex round. If I have ssDNA 25 random nucleiotids flanking by to primer biding site so totally 66mers. My targets protein will be MLL1. So I am wondering what are the factors to be considered regarding binding ratio of initial ssDNA library with protein and subsequent rounds? What should be the recommended binding ratio for the first and the following selex round?-Khun

Dear Khun,

This is something you should determine experimentally for your protein of interest and your library.Our biologists have provided a few publications which describes a RNA and DNA selection protocol which may have additional information

Design, Synthesis, and AmpliÞcation of UNIT 9.2 DNA Pools for In Vitro SelectionIn Vitro Selection of RNA Aptamers to a UNIT 24.3 Protein Target by Filter Immobilization

Best Regards,

Tiffany Teng

Product Specialist I

If 10ul RNA was used for 3′ adapter ligation, would you suggest using 10ul in 5′ adapter ligation reaction or the whole volume (20ul) from 1st reaction would work too?-Anastasia

Dear Anastasia,

Thank you for your email. While we recommend sticking to 2 uL of RNA input when possible, we found that we can still obtain good library prep results with up to 10 uL of RNA (without changing any other volumes in the 3′ Adapter Ligation Reaction). We recommend that you carry the entire volume of this reaction (10 uL + excess) to each of the next steps. This means that each subsequent reaction will now contain larger volumes than outlined in the manual.

Please let me know if you have any additional questions.

Regards,

Brea

My NTP shipment arrived without dry ice. Should I be concerned?

We ship our triphosphates on dry ice as a precaution. They are stable at room temperature, and even higher temperatures, for short periods of time. The chance of degradation during transit is very low. We recommend analyzing the material by AX-HPLC and then testing it in your application, provided the cost of the assay is not prohibitive. Contact us to request our analysis method.

Why is there a difference in pricing for molecular beacons depending on whether I have a license or not?

Our license to the technology requires that we pay a royalty for molecular beacons sold to institutions that do not have a license from The Public Health Research Institute of New York (PHRI). If you have a license, we do not have to pay a royalty to PHRI on your beacon. When you place your first order for a molecular beacon, we will ask if you have a license from PHRI.

How do you release products for shipping?

The batch record for your product is first reviewed and signed by the production manager in charge of manufacturing the product. All products are reviewed by a quality assurance specialist who signs the certificate of analysis verifying that all appropriate quality systems were followed and that all specifications were met prior to shipment.

Is there a list of restricted countries to which you cannot ship?

Shipping restrictions vary for specific countries and items. We recommend using a forwarding service for all shipments whose destination is not located in North America, Europe or Australia. This is mandatory if the shipment requires dry ice. We recommend using Biocair forwarding service; more information is available at https://www.biocair.com/Please inquire about your particular location.

How much does shipping cost?

Shipping costs vary depending on your location, whether or not dry ice is required, as well as the size of the dry ice box. All products delivered in solution will be shipped on dry ice. We ship via FedEx Priority Overnight or FedEx International Priority. Within the United States, standard shipping may range from $35-$350. International shipments have a greater range due to distance. If your order requires dry ice or insurance, additional charges will apply. Please inquire for pricing specific to your location and order.

Why is your turn around time longer than I expected?

The simple answer is that it takes longer to make a high quality, modified oligonucleotides reproducibly. TriLink was formed in 1996 largely in response to the poor quality of modified oligonucleotides available from many vendors. We developed very methodical protocols geared to the manufacture of highly modified products. Our protocols require a great deal of attention to detail, as much of the work must be done manually to ensure success. Additionally, our purification methods are very time consuming, and we include 1-2 days for the analysis of products prior to shipping.

How will my order be shipped?

We ship via FedEx Priority which guarantees receipt by 10:30 am the following business day to most locations within the US. Our oligonucleotides are packaged in microcentrifuge tubes as lyophilized solids. Oligos are most stable lyophilized; however, we can ship them in solution in water or buffer upon request (an additional charge may apply). Unless requested, all oligos are shipped at room temperature. They are stable at room temperature, but should be stored at –20°C upon arrival. Compounds in solution, with a dimethoxytrityl group must be shipped on dry ice to ensure stability of the trityl group. Please note: dye labeled oligos are light sensitive. All oligos are shipped in an inner reflective Mylar envelope to shield them from light. This envelope can be used for storage.

Can I speed up the delivery of my order?

Orders are prepared on a “first come, first served” principal. We do our best to accommodate your wishes by streamlining the process, however, our processes are generally firmly set, by both instrumentation and the training of the staff. Be aware that changing protocols may compromise the quality of the product. Most nights are already used for longer steps, such as deprotections, or dry-downs of larger samples. We can often schedule overtime if your requirements are truly urgent and you are willing to pay the added costs. This can save no more than a couple of days, since delivery is usually within 2 weeks. However, please discuss this option with us if you are interested.

How can I get a discount?

You can get a discount by signing an Annual Purchase Agreement (APA). Based on your forecasted order volume, you will be eligible for a discount for the following twelve months. Some products are already heavily discounted and are excluded from the Annual Purchase Agreement, and custom compounds are quoted on a case-by-case basis at the best price currently possible. Bulk discounts are also available for many products. Call or email for details.

Can I make changes to my order once it has been placed?

NTPs are shipped the following business day, unless otherwise stated. Changes or additions may be made to your order until the time of shipping, which is 3pm PST. For oligonucleotide or custom compound orders, if we can pull the order out of production before synthesis is started, we can accommodate your request without incurring an extra fee. However, once synthesis of the product has begun, changes to the sequence or scale are no longer possible unless costs of the current synthesis are met. Please keep in mind that you may be responsible for the cost of any specialized reagents that were purchased to synthesize your compound, plus a 10% surcharge. PAGE or HPLC purification may be added anytime before an oligonucleotide is shipped.

Which level of GMP do I need?

This is a decision for your regulatory department. However, we can offer advice based on your specific circumstances if you are unsure. Often, you do not need as stringent a manufacturing system as you may first believe.

Can I get research grade material that is well documented, but not as expensive as GMP grade material?

Yes! We find the costs of managing separate documentation systems (one for research grade material and one for GMP grade material) are more than the cost savings associated with the reduced documentation required for research grade materials. Therefore, at TriLink there’s no such thing as research grade material. All compounds are manufactured under a GMP system at comparable prices to research grade material from our competitors.

What do GMP, ISO and QSR mean, and how do they differ?

GMP stands for Good Manufacturing Practices, and refers to a system of manufacturing that guarantees reproducibility of product quality to set specifications. cGMP is simply Current Good Manufacturing Practices and refers to compliance with current regulations. It can be considered redundant since to be GMP compliant you must comply with current GMP regulations anyways. The requirements differ depending on what type of product is being manufactured and whether it is for pharmaceutical or diagnostic purposes. ISO stands for International Organization for Standardization, which offers a standard for operating a firm from management through manufacturing. It is more encompassing than GMP. QSR stands for Quality Systems Regulation, which are GMP standards described by the FDA for the manufacture of products for the diagnostic industry. The ISO and QSR systems each describe specific GMP standards. The ISO system pays more attention to the management of the firm and places a number of reporting loops in the firm to ensure attention to issues. The QSR system is more focused on the manufacturing systems, and the validation of those systems. Although neither standard is required to maintain GMP facilities, it is essential that a firm satisfies the requirements of the client’s quality system. Generally, this means conforming to ISO or QSR standards. Every product made at TriLink is manufactured under GMP. We are compliant with ISO 9001.

Where are you located?

We are located in San Diego, CA, in the Sorrento Mesa area just east of I-805 and just north of Miramar Marine Air Station. Our address is 9955 Mesa Rim Rd. San Diego, CA, 92121. Map It

Oops! How do I go back and add a modification in the middle of my sequence – it keeps inserting at the end?

The Internal Modifications click menu works by inserting the modification at the current end of the oligonucleotide you are building. If you need to go back and insert an internal modification after you have filled in your entire sequence, you will either need to:
1. Delete back to the position of the modification, select it in the menu and fill in your remaining sequence, or
2. Select the modification, then carefully cut and paste the modification into the correct position.
Please always be sure to double check your sequence before placing your order.

How do I insert a dye or conjugate on an internal linker?

Several modifications are available in the amidite form and can be added to your sequence simply by clicking on them. These modifications are marked “Amidite.” For all other internal dyes and conjugates you will need to choose the appropriate selective placement linker. To add an internal dye or conjugate click the linker of choice, followed by the dye or conjugate of choice from the internal modifications menu. The modification will appear at the end of your current sequence. Continue adding bases as needed. To view all available selective placement linkers, dyes and other non-fluorescent conjugates, see our Custom Oligonucleotide Components menu.

Can I order an oligonucleotide with a modification I do not see in the list?

TriLink offers numerous modifications that do not have standard list pricing and therefore do not appear in OligoBuilder®. These include 2’ Fluoro RNA bases, symmetrical branchers, cis-syn thymidine dimers, trimers and many more. Please email sales@trilinkbiotech.com for a quotation on any oligonucleotide modifications you do not see in OligoBuilder®. Be sure to include the sequence, backbone, modifications, starting synthesis scale or final yield and purification requirements.

Should I use the CleanAmp™ 7-deaza-dGTP Mix or just CleanAmp™ 7-deaza-dGTP alone?

Should I use the CleanAmp™ 7-deaza-dGTP Mix or just CleanAmp™ 7-deaza-dGTP alone?

For targets with <70% GC-content, addition of the CleanAmp™ 7-deaza-dGTP with standard dNTPs should amplify the target well. For targets with >70% GC-content, we recommend the CleanAmp™ 7-deaza-dGTP Mix for a robust amplification. Learn more about robust GC-rich target amplification in our white paper.

Have a question about CleanAmp™? Post a comment below.

How long does it take to activate CleanAmp™ dNTPs at 95°C and 60°C?

How long does it take to activate CleanAmp™ dNTPs at 95°C and 60°C? – Jia Wang

Thank you for your interest in TriLink. CleanAmp™ dNTPs have a t 1/2 of 40 minutes at 95°C. We are currently developing a new, improved version of CleanAmp™ dNTPs that have a t 1/2 of 7 minutes. We are also working towards an isothermal solution… Read More.

Have a question about CleanAmp™ dNTPs? Post a comment below.

Have a Question?

Collectively the TriLink Team has many years of experience in nucleic acid chemistry to offer. Post your question in the comment field below and we will get an answer to you as soon as we can.

Ask An Expert!

Welcome to our new Ask An Expert blog.  Answers to frequently asked questions  are listed by topic on the Ask An Expert homepage.  Please feel free to ask an expert by clicking on the comment link below. Thank you!