Enhanced Diagnostic Tools
Purification is recommended for all oligos greater than 40 bases in length and for many modified oligos. For demanding applications such as multiplex PCR, cloning, mutagenesis or antisense/RNAi methods, additional purification will significantly improve oligonucleotide performance. Every purified oligo up to 60 bases in length receives QC by capillary electrophoresis (CE) to document final purity.
Size exclusion desalting process has been used on all Bio-Synthesis's oligos. For oligo >10 mers
This process is based on a specific Reverse-Phase purification by using the difference in hydrophobicity between the full-length product (with a 5' DMT group) and truncated sequences (without DMT groups). Bio-Synthesis recommends this purification for all applications where full-length or high reproducibility is essential, including: cloning, PCR, mutagenesis, probes. The absence of salts and ammonium contamination, in addition to the absence of acid residues - which may occur after HPLC purification - make the cartridge purified oligonucleotides particularly suitable for use in cell culture. Two cartridge types:
The RP-cartridge for oligos from 8 to 39 bases long
The RP-LP cartridge for oligos from 40 to 80 bases long.
PAGE obtains extremely high purity, especially for unmodified oligos. This purification is based on molecular weight and charge, denaturing acrylamide gel electrophoresis provides a higher degree of purity than HPLC (93-95%). PAGE purification is strongly recommended for oligos over 60 bases in length. For unmodified oligos, purity of >90% is routinely achieved.
For the most demanding applications for high purity oligonucleotide, Bio-synthesis offers
Originally developed for isolating RNA oligos, RNase-Free HPLC Purification has been extended to DNA oligos for use when there is a high sensitivity to ribonucleases. RNase-Free HPLC purification is carefully performed in an RNase-free environment; reagents, equipment and lab surfaces are monitored for RNase contamination.
Bio-Synthesis's In Vivo Ready siRNA option provides high quality oligo for gene knockdown that are ready for introduction into animals. Each siRNA strand is HPLC purified, dialyzed to ensure that the siRNA is salt free (conductance of <1 microsiemens for a 50 µM solution), sterile filtered, and endotoxin tested. Of course, each siRNA strand is also analyzed by MALDI-TOF mass spectrometry and analytical HPLC, and the efficiency of annealing is verified by PAGE.
Guide for selecting purification grade
*Synthesis scale is not equal to guaranteed yield. Quantity varied depend on length and composition of the sequence, modifications and labeling.
Large RNA synthesis scale from mg to gram size are available, please call for quotation.
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