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DNA Oligos for Next Generation Sequencing Applications
Features and Benefits
- Optimized production process to ensure suitable purity and low levels of cross contamination.
- Stringent quality control (online trityl monitoring and MALDI-TOF MS or analytical PAGE)
- NGS oligos: ≤ 2 d
Trained scientists with molecular biology background are happy to support you (from 8 am to 5 pm)!
New and easy-to-use online portal with a series of helpful tools (e.g. order tracking & history, convenient search and re-order option
- Competitive pricing for volume as well as repetitive orders
- High guaranteed yields and/or average delivery yields
- Availability of current modifications (5’-Phosphate, PTO, 5-Me-dC etc.)
- Application independent of the instrument or technology
- Analytical HPLC available
- Certificate of Analysis available
- Various delivery formats possible (dried, liquid, tubes, 96-well plates etc.)
- Possibility to outsource the entire NGS project (from isolation to bioinformatics)
|Synthesis Scale||Purification||Length Restriction||Guaranteed Yield||Production Time [wd]|
|0.04/0.2 umol||HPLC||13 – 65||3 OD||≤ 2|
|PAGE||66 - 99||1 OD||≤ 3|
|1.0 umol||HPLC||13 – 65||15 OD||≤ 2|
|PAGE||66 - 99||7 OD||≤ 3|
It is crucial to emphasize the importance of oligonucleotide purity for NGS applications. Most researchers take into account the n-x side products when they consider oligonucleotide purity, but do not ask their oligo suppliers for post-synthetic processes that have been optimized in terms of oligo cross-contamination.
Therefore many suppliers use cheaper high-throughput methods (e.g. reversed-phase cartridge purification) to purify NGS oligos, which however are especially prone to cross-contamination. As a consequence, even a small amount of cross-contamination which is harmless in less sensitive applications (e.g. PCR or Sanger), may become a severe problem in the NGS data analysis since incorrect amplification products are likely to be sequenced as well. A typical example is the risk of barcode misinterpretation in multiplexing experiments where index adapters containing the MID (multiplex identifier) or barcode are used.
Consequently, Microsynth has established and validated a specific process for production of NGS oligos that is based on either stringent HPLC or PAGE purification procedure where the elimination of cross-contamination is given the highest priority.
How to Order
Option 1: When ordering NGS primers, please proceed as follows:
- Enter our webshop, click on DNA in the blue “DNA/RNA Synthesis” domain
- Select either Normal Entry in order to type or copy/paste the desired sequence information etc. or alternatively select Upload Entry by using our convenient Excel Template (can be downloaded during ordering)
Important: You must use the prefix “NGS_” ahead of your oligo name (e.g. NGS_Cart_fw01), select either 0.2 µmol or 1.0 µmol scale and further choose HPLC purification (up to 65 mers) or PAGE purification (66 to x mers)
Follow the further instructions
Option 2: When ordering primers for Illumina amplicon sequencing, please proceed as follows:
- Use the dedicated “Order Form_Illumina_AmpliconDeepSeq” (can be downloaded in the right column); in this context see also our application note “Amplicon Deep Sequencing”)
- Specify now the locus-specific sequences for your first-step PCR primers and then select your desired indexed forward and reverse primers
- Enter our webshop, click on DNA in the blue “DNA/RNA Synthesis” domain and then on Upload Entry
Follow the further instructions