Reduce barcode crosstalk in your multiplexed NGS experiments
Quality requirements for synthetic Oligos used in Next-generation sequencing (NGS), such as indexed adapters and barcoded fusion primers, have reached unprecedented high levels. Traditional purification methods (e.g., HPLC) can effectively increase the number of full-length Oligos but are incapable of reducing cross-contamination that can occur in co-synthesized Oligos. In contrast to less sensitive applications where such Oligo-to-Oligo crosstalk can go unnoticed, the use of indexed adapters containing even low levels of cross-contamination can lead to barcode misalignment and sample misidentification, which may confound experiments requiring detection of rare variants. NGS Grade Oligos are manufactured by proprietary methods that are proven to reduce index cross-talk and increase the success of multiplex NGS experiments. With NGS Grade oligos, you can:
Our oligos come with a guarantee of high performance for a wide spectrum of research applications including:
Bioserve India provides the following methods for Purifying Oligonucleotides
All oligos synthesized are processed through normal phase chromatography column which removes salts but not failure sequences. Desalted oligos are ready-to-use, suitable for many PCR and sequencing applications without further purification.
Recommended Applications Like PCR, AFLP, RFLP
This purification method is based on the “trityl-on” synthesis of the oligonucleotides. Purification on reverse phase cartridge ensures higher purity scales. It removes failure sequences. As the length of oligos increases, the proportion of truncated sequences tends to increase. These impurities will not be removed by Cartridge and thus for longer oligos, HPLC or PAGE is recommended.
Recommended Applications: Like Sequencing, RFLP, DNA Fingerprinting, RT-PCR, First strand DNA synthesis. Cartridge purification is the recommended purification for Oligos greater than 35 to 50 bases.
BioServe India provides both reverse phase and ion exchange HPLC purification of oligonucleotides. HPLC is an efficient purification method for oligos with fluorophores, as their intrinsic lipophilicity provides excellent separation of product from its contaminants. RP-HPLC is a method of choice for larger scales due to the capacity and resolving properties of the column. As resolution depends on lipophilicity it will decrease with the length of the Oligo. RP-HPLC is usually not recommended for purifying products longer than 60 bases.
Recommended Applications like Antisense studies, Cloning adapters, Real Time PCR.
In this procedure, oligonucleotides are purified based on their mobility on denaturing polyacrylamide gels. The band of interest is excised using UV shadowing and extracted from the gel. Oligonucleotides purified by this method are 99% full length. PAGE is the recommended purification for longer oligos (>50 bases).
Recommended Applications like Mutagenesis, Gene Synthesis, Primer Extension, Cloning adapters.
Committed Guaranteed yield on synthesis parameters
All the oligonucleotides synthesized at BioServe-India are quantitated using the absorbance at A260.
All the oligonucleotides synthesized are electrophoresed on a polyacrylamide gel to make sure that they are of the requested length and there are no n-1 oligonucleotides. This gel picture will be provided to the customer.
Each shipment includes data sheets for import into customers’ information systems containing well location, sequence, oligo name, lot number, MW, µg/OD, length, Tm, concentration and volume (in solution).