BioMicroCenter:Oligo Synthesis

From OpenWetWare
Jump to navigationJump to search

HOME -- SEQUENCING -- LIBRARY PREP -- HIGH-THROUGHPUT -- COMPUTING -- OTHER TECHNOLOGY

** All users must be trained before being allowed to use the equipment **

The BioMicro Center currently hosts two oligo synthesizers: one Dr Oligo 96 (Biolytic) and one Syntax STX-200 (DNAScript).

Dr Oligo is optimized for higher concentrations of oligos made through polyamidite synthesis. For ready to use oligos a series of steps must be completed using the other related machines in the Center including the column presser, the heated pressure chamber and the processor: each individual step utilizing a machine takes a few hours of setup, processing and completion. These tools are available to MIT researchers for use in their research and may be signed out using iLabs after training is complete.

The Syntax utilizes enzymatic chemistry to create a set of 96 phosphorylated ready to go oligos utilizing the platform solely in about a day. Plates may be ordered by emailing biomicro@mit.edu.

Dr Oligo 96

Instrument Dr Oligo 96
USAGE Walkup - MIT only
Approval Required
UNIT Per hour + Consumables
Number of oligos per run
  • 24
  • 96
Hazards
  • Pressurized Synthesis
  • Heated Pressure Chamber with Ammonium
SCHEDULE USAGE ilabs
NEW USERS New users should request training by emailing the BioMicro Center team.
DONATED BY Dr Mark Bathe, CEHS


"Dr Oligo" is the BioMicro Center oligo synthesizer optimized for high concentration. A 24 or 96 column plate can be loaded on the synthesizer--it is recommended to duplicate oligo sequences and not necessarily use dummy columns since oligo synthesis can fail for some columns during polyamidite synthesis. This synthesis adds 3' to 5', and each subsequent base addition reduces yield. Synthesis of oligos lower than 50 bp on this platform is typically short and high yield, oligos longer than 100 or 150 bp have a higher likelihood of failure. Post processing, size selection is recommended since shorter non-ideal oligos may also be eluted.

Users should plan synthesis in advance to coordinate usage with other users.


synth process

heat

STX-200

Instrument Syntax STX-200
USAGE Order through BMC
UNIT Per hour + Consumables
Chemistry
  • Enzymatic
  • 5' phosphorylated end
Number of Oligos per Run
  • 96 only
ON LOAN THROUGH DNAScript


"The Syntax" platform employs enzymatic DNA synthesis. All chemistry is handled within the Syntax machine-final elution of the oligos occurs in a 96 well deep well plate. Final concentration is at least 10x lower than mid-yield Dr Oligo products.


inline