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For easy ordering, check out out the Invitrogen Custom DNA oligos page.
Invitrogen Custom DNA oligos are synthetic oligonucleotides made according to your specifications and can be used in a variety of applications, from PCR and sequencing to probes for gene detection. Custom oligos are offered with standard deoxynucleotides, modified bases, and 5´- and 3´-modified nucleotides. Available modifications include fluorescent dyes, enzyme conjugates, and S-oligos for antisense studies. We offer five standard synthesis scales and four purity options (Table 1).
Oligos are made using a DNA synthesizer, a computer-controlled reagent delivery system. DNA is synthesized in the 3’ ⇒ 5’ direction, and each base addition is accomplished through a series of chemical reactions. But no chemical reaction is 100% efficient; during DNA synthesis, the maximum coupling efficiency obtainable is normally around 99%. Therefore, approximately 1% of the growing nucleotide chains fail to couple to a base in each round. In the case of a 30-mer oligo synthesis, the final product would also contain 29-mer failures, 28-mer failures, etc. For an oligo of this length, the percentage of full-length oligo is estimated to be between 74% and 54%, assuming a 99% or 98% reaction efficiency. The percentage of full-length oligo produced decreases as the length of the oligo increases.
The failure sequences inherent in the oligo synthesis process can compete with the full-length product in some applications; therefore, to help ensure success in downstream experiments, it may be necessary to purify the full-length oligo away from the failure sequences.
Purification method | Description | Benefit |
---|---|---|
Desalt | Oligos are processed through normal phase chromatography columns which removes salts but not failure sequences | A salt-free DNA solution, ready to use; suitable for many PCR and sequencing applications without further purification |
Cartridge | Based on reversed-phase chromatography; removes failure sequences from the completed synthesis | Provides full-length sequences needed in some applications |
HPLC | Reversed-phase high-performance liquid chromatography (HPLC) removes failure sequences or unincorporated label the same way as cartridge purification | Guarantees highly purified primer required in some applications (≥85% full-length) |
PAGE | Method used to differentiate full-length product from failure sequences based on size and conformation | Provides the highest percentage of full-length oligos (≥85%) required for certain demanding applications such as mutagenesis or adapter production |
For easy ordering, check out out the Custom DNA oligos page.
仅供科研使用,不可用于诊断目的。