MGH   
CCIB
 
96-Well Plasmid DNA Preparation

 

Our Laboratory Automation group has developed a purification system to isolate high-quality plasmid DNA in 96-well format. The purified plasmid DNA is suitable for a broad range of downstream applications, including transfection, in vitro expression, automated fluorescent DNA sequencing and cloning.

We currently offer our plate-based DNA preparation service either as a stand-alone service or in combination with our DNA sequencing options.

Service Description:

  • User will submit 96-well deep well culture block with cell pellets.
  • Bacterial cells undergo alkaline lyis, and the released DNA is bound to an in-house developed spin filter. The plasmid DNA is eluted in 50 ul 5 mM Tris-HCl, pH 8.5. A small aliquot of purified plasmid DNA is used for quantitation with a SpectraMax Plus 384 plate reader, and another aliquot will be removed for sequencing (if ordered). The remaining volume is approximately 30 ul (per well).
  • Using the Janus Mini MDT Automated Workstation in combination with a low-speed benchtop centrifuge, we can process up to four 96-well plates simultaneously.
  • DNA yield might vary depending on plasmid copy number, E. coli strain, and growth conditions (growth medium, incubation time, aeration).
  • Turnaround time depends on the total number of plates received but is typically 1-2 business days. The same turnaround time applies for a combined 96-Well Plasmid Preparation PLUS Sequencing request. Sequencing data are usually available within 24 hours upon completion of the plasmid preparation order.
  • Upon completion of an order, clients will receive an automated email stating that their order is ready to be delivered to the desired remote drop box or can be picked up at the core facility; the quantitation data (OD260, OD280, 260/280, DNA concentration) will be included as an attachment.
  • Plasmid DNA preparations are returned in a sealed V-bottom polystyrene plate and will be delivered to the appropriate remote drop-box location via our free Courier service (currently for internal customers only). If preferred, orders can also be picked up directly at our facility.

Please note: We recommend that you retrieve your plasmid DNA preparations as soon as possible. We will discard preps after approximately four weeks if they have not been picked up.

To ensure efficient processing and quality results, please review our requirements for sample submission listed under the Plasmid Prep Guidelines.