Choosing the right DNA oligos for your application

Oligonucleotides are the starting point for many of today’s biology research, drug discovery and diagnostics applications. These highly technical applications demand high-quality oligonucleotides for success. Custom DNA oligos at Life Technologies are synthesized on a highly automated, computer-controlled system using standard cyano-ethyl phosphoramidite chemistry. Coupling efficiencies are monitored throughout synthesis of each oligonucleotide by trityl analysis, ensuring the quality of the process, not just the end product. Post-synthesis QC, mass spectrometry for short oligos and capillary electrophoresis (CE) for long oligos, ensure quality.

Choosing the synthesis scale and purification option that is best for your application depends on the nature of your downstream applications. The table below is designed to help you choose the right oligos and purification methods for your application.

In general, Cartridge purification is fast and economical but may compromise purify or yield depending on the oligo sequence (sequences with 5’ G’s can be problematic). HPLC is more time consuming but gives excellent purity up to 55 bases. Above 55 bases, it becomes increasingly difficult to differentiate full-length product from n-1 failure sequences. PAGE gives excellent resolution even with very long oligos but generally sacrifices yield for quality.

 

Application

Desalted

Cartridge

HPLC

PAGE

 

  • 25 nmole – 10 µmole
  • 5 – 100 bp
  • Removes salts but not failure
  • 50 nmole – 1 µmole
  • 7 – 55 bp
  • Removes failure sequences
  • Provides 80% full-length sequence
  • 50 nmole – 10 µmole
  • 10 – 55 bp
  • Removes failure sequences
  • Provides >85% full-length sequence
  • 50 nmole – 10 µmole
  • 7 – 100 bp
  • Removes failure sequences
  • Provides > 90% full-length sequence

Standard PCR

  

 

Specialty PCR

  

Cloning

  

cDNA Library Construction

  

Sequencing

  

 

Fluorescent Sequencing

 

 

 

Next-Generation Sequencing

  

Mutagenesis

  

Antisense

  

 

Gel Shift Assays

 

 

Microarrays