96-Well Plasmid DNA Preparation: FAQs
I would like to use your 96-Well Plasmid Preparation service. How do I set up a new account?
To create a new account, please click the Create New Account link (to be found on the top of all the pages of our website) and fill out the required fields of the Registration Form. To complete the registration process, click on the Register My Account link. Your account will be active immediately after registration, and you are ready to submit your first order!
For what applications is the purified plasmid DNA suitable?
Our purification system consistently provides high-quality plasmid DNA suitable for a broad range of downstream applications, including transfection, in vitro expression, automated fluorescent DNA sequencing and cloning.
Which growth medium should I use?
Our 96-well Plasmid DNA Preparation procedure has been optimized for the use of Terrific Broth (TB). The use of a rich medium such as TB requires the optimization of the growth time.
What is the recommended optical density for the bacterial cultures?
Our 96-well Plasmid Preparation protocol is optimized for use with bacterial cultures grown to an OD600 value of 2.5-2.8. Densities higher than 3.0 may cause our proprietary filters to clog and the prep may fail.
What type of seal do you recommend for plates/blocks?
A proper seal is very important to avoid leaks and cross contamination. We recommend the use of aluminum seals from Worldwide Medical Products, Inc. (Aluma Seal II, 100/bx; Cat # 41061019). Although a plastic seal may seem sufficient, we have found that they often loosen during transportation.
How should I label my 96-well block?
Please label the front of your growth block as clearly as possible with the Automation order number (label faces you when well A1 is located in the top left-hand corner). If you wish to have your purified plasmid DNAs sequenced at our facility, please label the tube containing the appropriate sequencing primer with the Automation order number as well.
What type of buffer will be used to elute my samples?
All samples are eluted in 5 mM Tris-HCl, pH 8.5. Contact us or clearly make a note on your Automation submission sheet if you would like your samples eluted in sterile nuclease-free water.
What kind of yield can I expect from your 96-well Plasmid Purification protocol?
Yield might vary depending on plasmid copy number, E. coli host, and growth conditions (growth medium, incubation time, aeration). For high-copy-number plasmids, yields up to 5-7 µg can be expected if optimal growth conditions and host strain have been selected.
Can my purified plasmid DNAs be sequenced directly at your core facility?
Yes. We currently offer our plate-based DNA preparation service either as a stand-alone service or
in combination with our DNA sequencing options.
If you would like your purified samples to be sequenced at our facility, please request the
96-Well Plasmid DNA Preparation PLUS Sequencing option when ordering.
Provide us with a copy of your submission sheet(s) along with a separate tube containing 400
µl of the appropriate sequencing primer (provided at a concentration of 10ng/µl
(1.6 µM)). Please note that well H12 MUST be left empty for an internal quality control!
If you are planning to submit multiple sequencing primers with your plate of harvested bacterial cells, please array your primer set in a separate 96-well PCR plate (at least 10 ul per well, at a concentration of 10 ng/ul (1.6 uM)). Well positions of primers must match the well positions of your plasmid clones.
If your DNA templates should contain GC-rich regions, hairpin structures,
or repeats (di- and trinucleotide stretches), we highly recommend that you order
the Difficult Template Sequencing service to ensure successful sequencing.
What is the turnaround time for 96-well Plasmid Purification order?
Turnaround time depends on the total number of plates received but is typically 2 business days. Upon completion of an order, clients will receive an automated email stating that their order is ready to be returned to the appropriate remote drop-box or that it can be picked up at our core facility.
If you have requested our combined Plasmid Prep PLUS Sequencing service, your purified plasmid DNAs will be returned as soon as the sequencing data is available.
How will my purified samples be returned to me?
Upon completion of an order, you will receive an automated email stating that your order
is ready to be returned. The quantitation data
(OD260, OD280, 260/280, DNA concentration)
for your order will be included as an email attachment. Your purified plasmid DNA samples
(packaged in a sealed v-bottom polystyrene plate, with ~30 µl DNA solution per well)
will be delivered to the same remote drop-off location
where the original block has been
picked by our courier service (currently for internal customers only).
For a detailed list of our remote pick-up locations, please see here.
Contact us or leave a note with your submission sheet if you wish to pick-up your samples directly at the core 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.
Can I submit samples during weekends/holidays?
Unfortunately not. The CCIB DNA Core operates at a reduced capacity over the weekend and is closed on Sundays and MGH holidays. The courier service is not available on weekends and on holidays observed by MGH.