Custom Plate Oligonucleotide Synthesis
Oligonucleotide synthesis is an extremely useful technique in many experiments. It can produce relatively short nucleic acid fragments with a defined chemical structure (sequence) through chemical synthesis. Creative Biolabs has designed and built a standard 96-well plate-based phosphoramidite chemical reaction scheme that can synthesize standard, degenerate or modified oligomers on a single plate. These oligonucleotides can be used as probes to detect complementary DNA or RNA by molecular hybridization, as tools to target the introduction of mutations and restriction sites, and to synthesize artificial genes.
Oligonucleotide Synthesis
The reactivity of naturally occurred nucleotides (nucleoside 3'- or 5'-phosphate) and their phosphodiester analogs is generally insufficient to provide rapid synthetic preparation of high yield oligonucleotides. However, if the 3'-O-(N, N-diisopropyl phosphoramide) derivatives of nucleoside (nucleoside phosphoramide) can be used as the basic material of phosphite triester method, the selectivity and speed of internuclear bond formation can be greatly improved. In solid-phase synthesis, the assembled oligonucleotide is covalently bound to a solid support through its 3'-terminal hydroxyl group and remains attached throughout the entire process of strand assembly. The solid support is included in the column, and the size of the column depends on the scale of synthesis and may vary from 0.05 ml to several liters. Recently, high-throughput oligonucleotide synthesis, in which solid support is contained in the wells of a multiwell plate (typically 96 or 384 wells per plate), has become an option for small-scale parallel synthesis of oligonucleotides. At the end of the strand assembly, the oligonucleotide is released from the solid support and eluted from the column or well.
Figure 1. Plate oligonucleotide synthesis chamber schematic. (Rayner, 1998)
Custom Plate Oligonucleotide Synthesis
Our plate oligo synthesis service includes the following parts:
- Chemical synthesis of oligonucleotides with different specifications. The quality of the oligonucleotide was evaluated by a combination of capillary electrophoresis (CE) and HPLC, and the reaction parameters were appropriately adjusted based on the feedback results.
- Postprocessing and analysis after oligonucleotide synthesis. The mass spectrum of the oligonucleotide was measured using mass spectrometry. The absorbance at 260, 280 and 320 nm was taken to measure the amount of product.
Advantages
- Single base synthesis cost is low, which can save your cost when you need a large number of synthesis.
- The procedures are designed to be used in a high throughput environment and require a minimum amount of user intervention.
- Through two-dimensional operation, the design also has the capacity to increase the scale of the synthesis by ensuring sufficient space in the external chamber to accommodate four reaction plates and a second injection head.
- The design of the instrument and its operating software allow using different synthesis formats such as 48-well, 96-well and 384-well plates to accommodate different synthesis scales.
If you have any questions about our custom plate oligonucleotide synthesis service, you can contact us by email or send us an inquiry to find a complete solution.
Reference
- Rayner, S.; et al. (1998). MerMade: an oligodeoxyribonucleotide synthesizer for high throughput oligonucleotide production in dual 96-well plates. Genome research. 8(7): 741-747.
