PLASMID DNA PREPARATION

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High Copy Plasmid Prep: 96-well block format only.  

$175/block for UC labs. 

We sell the culture blocks at our Facility, along with gas permeable membrane covers for $4.50. 

We only accept "Costar 3961" culture blocks, as they are compatible with our instrument AND our protocol. 

Please ensure you use the correct blocks! If your samples are in a different block, please contact us to discuss your options for robotic processing.

This service utilizes magnetic bead technology for high-throughput purification of plasmid DNA from E. coli. It works well for high copy plasmids.  For low copy plasmids, please see the special instruction below for growing the cultures.

If you have the ability to pellet your cells and remove the media, that is helpful (to us). You can then seal and freeze the block for future processing at our Facility. 

We do not accept culture blocks on the weekend, but if you have access you can leave them in the cold room on the 3rd Floor, WITH YOUR PAPERWORK... and please email us to let us know they are there. 

To take advantage of this service, you will need:

1. 96-well 2.2 ml deep well culture block with gas permeable seal (pick these up in 310 Barker)
2. Up to 95 samples (please reserve well A1 for our sequencing control).
3. Cultures grown in 1~1.5mL of 2YT broth or Terrific broth with antibiotic in deep well culture block. 
4. Start from either a single colony or a glycerol stock, and culture with shaking at 37°C to mid or late log phase (16-20 hours).
5. Please bring your prepared culture block with your filled-in  Plasmid Prep Order Form to 310 Barker Hall.
6. Once the plasmid preparation is complete, we can sequence your plasmids for you.  For Sanger sequencing -- please clearly indicate which stock primer(s) you would like us to use, and/or provide sufficient amount of your own custom primer at a 10μM concentration when you drop off the culture block - providing 1µL per sample. ALTERNATIVELY -- if you would like full-plasmid Nanopore sequencing, we can do that too. For either sequencing case, please provide the filled-in sequencing form, and we will do the rest!
7. Please email a copy of  the dowloaded Full Plate Sequencing Form to dnaseq@berkeley.edu

Generally speaking it is advantageous to grow cultures for 16-20 hours (possibly even longer) to improve yield. Paricularly for Low Copy Plasmids, please grow your culture in culture tubes using Terrific Broth or 2YT broth + antibiotic (AVOID PLAIN OLD LB Broth) with shaking  for 18-24 hours, and transfer 2ml of each culture into one well of the 96-well block.