| Column | Capacity |
|---|---|
| DNA-99-3510 | 2 µg |
| RPC-99-3810 | 50 µg |
| RPC-99-2110 | 600 µg |
| RPC-99-3015 | 2 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.
Can I get single base-pair resolution in my separations?
This will depend on the length and complexity of your sequence. For very short fragments, single-base resolution is possible. For longer lengths, (hundreds of base pair) single-based resolution is not possible, but tens of base-pairs is resolved. For longer yet, resolution of adjacent peaks is only at hundreds of bases.
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.
How do I prepare my column for storage?
The column needs to be flushed with Solution D for at least 30 minutes. The column should then stay filled with Solution D when stored.
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.
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 I load IVT product directly to the columns?
Yes, RNA from IVT reactions can be purified on our columns without preliminary purification.
What is the RNA ladder used to generate chromatograms?
We used the RiboRuler High Range Ladder from Invitrogen.
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 |
To learn more about the HPLC columns ADS Biotec offers check out our Nucleic Acid HPLC Columns