| Column | Capacity |
|---|---|
| DNA-99-3510 | 2 µg |
| RPC-99-3810 | 50 µg |
| RPC-99-2110 | 600 µg |
| RPC-99-3015 | 2 mg |
| UHC-99-3510 | ~1.7 mg |
| UHC-99-3810 | ~4.25 mg |
Is there a maximum sequence length that can be successfully separated using the columns?
No, there is no maximum as the mechanism of separation remains the same for small and large nucleic acids. However, the larger the sequence is, the longer the retention time will be, and the longer runs will take.
Which columns can separate single strand (ssRNA) and double strand RNA (dsRNA)?
Both ssRNA and dsRNA can be separated and purified on our RNASep columns. There will be differing retention times for the distinct species and separation can be conducted once retention times have been established.
Are there column heaters available for RPC-99-2110 and RNA-99-3015?
Our column products have different dimensions of diameter and length but are compatible with most HPLC systems’ column oven or heating devices. Column heaters are available at Timberline Instruments and are compatible with ADS Biotec columns.
Can all RNASep columns be used for separation and purification?
Yes. We offer a range of sizes to best fit your needs. All RNASep columns are packed with the same nonpolar material to separate and purify RNA.
Can all RNASep columns be used for poly-A tailed RNA analytical separations and purification?
Yes. The RNASep technology uses reverse phase ion pairing mobile phase chemistry reliant on the negative charge of nucleic acids and the positive charge of TEAA, so specific sequences with poly-A tails will blind the column.
Do I need to filter HPLC buffers from ADS Biotec?
No, HPLC buffers are filtered during manufacturing and do not need to be filtered at the point of use.
Will the RNASep column fit on my HPLC system?
ADS Biotec columns use an industry standard end-fitting 10-32 that makes them compatible with nearly all established HPLC Instruments. The exception would be chromatography systems geared for industrial scale that typically use very high eluent flows and lower pressure in their chromatography systems.
How long will the RNASep column last? How many injections are possible?
Some customers have had thousands of injections on a single column over multiple years. The life of the column in typically dependent on the cleanliness of the sample and the cleaning regimen used to wash the column post-analysis. Follow the column care guide to maximize the lifetime of the column. Typical life for analytical columns is usually much higher than large loadings made to the larger semi-preparative columns.
How do I prepare my column for storage?
The column needs to be flushed with 100% Solution D at 1.0mL/min for at least 30 minutes before removing for storage. The column must be stored in Solution D at room temperature.
What column is best for my needs?
RNA-99-3810 is best for separation, purification, and QC of RNA molecules. RPC-99-2110 is best for intermediate scale up. RPC-99-3015 is best for RNA purification for larger scale up.
What sort of entities have used your columns and/or buffers?
We have a wide range of clients that use our RNASep columns and separation buffers, in vaccine and therapeutic development, RNA purification and analysis in startups, as well as the established pharma, and biotechnology companies globally. They have been used for nucleic acid analysis in over 30 countries and for over 2 decades.
How long after placing an order can I expect to receive my order?
For the separation buffers, stock is kept on-hand and turnaround is rapid. Large scale or customer orders can take 8-12 weeks. RNASep columns are typically shipped within 15 days of being ordered.
How long has ADS Biotec been ISO 13485:2016 certified?
– ISO 9001 certification was achieved starting in 2008 and was maintained until December of 2023
– ADS Biotec obtained certification to the ISO 13485:2016 standard as of January 2023 and has maintained certification since.
How does PS-DVB compare to Silica gel?
The PS-DVB columns allow for increased separation efficiency and faster separations when compared to silica gel packed columns, with better ‘longevity’ and robustness in that they can be flushed and reused for more separations. Feedback we have received from customers indicates that our media is more consistent and robust after repeated usage compared to silica columns.
How long do I need to equilibrate the column when changing temperatures?
The column should be equilibrated with 1:1 Buffer A:Buffer B for a minimum of 30 minutes before preparing for sample injection.
What types of nucleic acids can be separated using ADS Biotec columns?
Any type of nucleic acids can be separated using our columns. Due to the mechanism of separation, the length of the species is not a factor. PS-DVB columns have been used to purify mRNA, cRNA, siRNA, asRNA, miRNA, etc.
What is the pore size on the macro-porous, high-capacity column media (NUC-99-3860, RPC-99-3870)?
The nominal pore size is estimated at 80 Angstroms.
Can ADS Biotec columns be used to purify 5 kb long sequences?
Yes, our columns are able to purify RNAs that are 5 kb in length.
Can the columns be loaded with in-vitro transcription (IVT) product without damaging the columns?
Yes, IVT can be safely loaded to the columns without any expected adverse effects. See DOI: 10.1016/j.omtn.2019.02.018 for published information regarding IVT on the columns.
What is the RNA ladder used to generate chromatograms?
We used the RiboRuler High Range Ladder from Invitrogen.
What pressure will the column tolerate?
Maximum pressures should not exceed 3000 psi (207 bar). Typical operating pressures are 600 psi to 2600 psi (41-179 bar).
What temperature will the column tolerate?
Maximum temperature is up to 100 deg C. Typical operating ranges are 45 to 75 deg C.
What pH will the column tolerate?
The columns will typically tolerate a wide range of pH, from pH 0 to pH 14.
What products should not be put on the columns?
– The following components should be avoided in PCR reactions prior to injection to the column: Bovine Serum Albumin (BSA), Autoclaved Water, Mineral Oil, Formamide, Proteinase K, enhancers, stabilizers, additives, Glycerol (>2%), DMSO (>10%), and Betaine- 1.25 (>2.5M)
– Addition of the following components will void the column warranty: High molecular weight stabilizers >1% (ex. Polyethylene glycol – PEG) and Detergents >1% (ex. Triton X-100, NP40, Tween 20, SDS).
What is the HS Code (HTS Code) for the RNA columns?
– For export: 9027.90.8950
– For import: 9027.90.5625.
To learn more about the HPLC columns ADS Biotec offers check out our Nucleic Acid HPLC Columns
How are the buffers stored and shipped?
Buffers are provided as a pre-mixed, ready to use solution in 2.5L bottles stored at room temperature protected from light and heat. Some buffers have high enough acetonitrile levels to be classified as dangerous goods. These flammable products are segregated to an explosion-proof room and smaller volumes are stored in flammable cabinets when not in use. Argon gas is used to sparge the liquid of entrained gas and is added to the headspace on bottles to improve long-term stability. Please carefully consult the Safety Data Sheets for storage and transport requirements of the buffers. Buffers can be shipped around the globe, mainly by sea freight.
How are buffers qualified before release?
A battery of tests is conducted on the final product, including actual testing in operating the WAVE HPLC system to verify the quality and performance are consistent.
Are there recommended buffers to be used with the RNASep columns?
We recommend the following for use with RNASep columns: ADS Biotec Cat. # 553421, 553422, and 553423.
Is your 2M TEAA Buffer RNase free?
Yes, we have determined that our 2M TEAA buffers are RNase free. Additionally, our source water has been tested and is RNase free.
What is the different between TEAA and HAA?
– HAA is HexylAmmonium Acetate. TEAA is Triethylammonium Acetate. Both are ion-pairing reagents that work well with Nucleic Acids.
– HAA improves the separation efficiency of challenging nucleic acid sequences. These sequences are often G-C rich, for which HAA can help to give better separation of differing lengths in the sample. HAA buffers typically used with RNAs and Oligonucleotides under 200 nt in length.
What is the HS Code (HTS Code) for the Ready-to-use HPLC Buffers (aka Eluents, aka Mobile Phases)?
– For export: 3822.00.0002
– For import: 3822.00.5095.
To learn more about the mRNA Purification Buffers ADS Biotec offers check out our Nucleic Acid HPLC Columns
What are the recommended flow rates and pressures for the columns?
| Column | Flow Rate (mL/min) | Pressure Range (psi) |
|---|---|---|
| DNA-99-3510 | 0.7 – 1.1 | 900 – 1600 |
| RPC-99-3810 | 0.75 – 1.5 | 800 – 1400 |
| RPC-99-2110 | 4 – 8 | 800 – 1400 |
| RPC-99-3015 | 4 – 10 | 800 – 1600 |