Rapid Plasmid Sequencing: Sample Submission
To ensure the successful sequencing and assembly of your samples, all guidelines for sample preparation and submission must be addressed. Please contact us if these requirements cannot be met so we can work with you on identifying a solution on a case-by-case basis.
Sample Requirements:
Please submit CIRCULAR DNA only! Linear DNA such as amplicons and restriction fragments have to be submitted for our Complete Amplicon Sequencing service, as our Rapid Plasmid Sequencing process is not compatible with linear DNA.
Plasmid size is limited to 25 kb. For larger plasmids please use our Complete Plasmid Sequencing service.
The purified plasmid DNA should be resuspended in nuclease-free water or 10 mM Tris-HCl (pH 7.5-8.5).
The plasmid DNA must be diluted to a final concentration of 25 ng/µl. Please note: The DNA concentration should be determined using a Qubit or similar fluorometric method. Nanodrop and other UV spectrophotometers do not yield reliable measurements since they do not specifically measure nucleic acids.
Our workflow requires the submission of 20 µl of the diluted plasmid DNA.
We strongly encourage you to submit a reference sequence for your plasmid if possible. A reference sequence represents your best estimate of the plasmid's sequence; it does not have to be perfect.
Send your samples in either PCR strip tubes or 96-well PCR plates.
PCR Strip Tube Orders:
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Please submit your samples in standard 0.2 mL PCR strip tubes. Please note: Our operational set-up does not accept 1.5 ml snap-cap microcentrifuge tubes and screw-cap tubes.
Please label the first tube of each strip with the last 3 digits of your order number. If submitting multiple tube strips, please always label the first tube with the last 3 digits of your order number and label the second tube with the number of the strip.
For example, for order number 229108n containing 17 samples the first strip of tubes (samples 1-12) would be labeled with 108 on the first tube and 1 on the second tube. The second strip of tubes (samples 13-17) would be labeled with 108 on the first tube and 2 on the second (please see photo).
PCR Plate Orders:
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Please use standard PCR plates. We recommend the use of semi-skirted or full-skirted plates to prevent the PCR plate from bending during transit, which could result in loosening of the seal and subsequent sample loss and cross-contamination.
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Please arrange your samples horizontally (in rows) in the PCR plate. Proceed from well A1 to A12, then B1 to B12, then C1 to C12, etc.
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We accept partial plates but please do not leave any empty wells between samples, and please do not leave any empty spaces between samples on the order form.
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The order form must match the order and position of samples in the PCR plate.
Please label the plate as clearly as possible with the complete order number.
Tightly seal your plate with an adhesive foil or strip caps to prevent sample loss or cross-contamination.
Sample Shipment:
Please submit your sample(s), including a hard copy of the completed order form, by using one of our remote dropboxes (please see here) or by delivering it personally to our core facility.
If remote or direct sample drop-off should not be an option for you, please ship your samples, including a hard copy of the completed order form, via a courier delivery service of your choice (for example, overnight shipment via FEDEX, UPS, etc.) to our core facility. Samples can be shipped at ambient temperature.
Shipping Tubes:
Unfortunately, we often receive sample tubes that were damaged during shipment (cracked or shattered tubes, sample loss due to leakage or evaporation). Therefore, we strongly recommend protecting them with bubble wrap or several layers of Kimwipe tissue within a secondary container such as a Falcon tube or small cardboard or plastic box.
Shipping 96-well Plates:
To prevent sample loss and/or cross contamination, we recommend tightly sealing all wells of the plate with strip tube caps. PCR Tube Strip Flat Caps (Eppendorf cat# 0030124847 or VWR cat# 75874-710) work well for most plates. Our everyday experience tells us that plate foils/plastic seals can partially lift off or even entirely detach during transit dependent on the shipping conditions (heat, air pressure, humidity, etc.). Please place your plate in a corrugated box for maximum protection.
Shipping Address:
MGH CCIB DNA Core Facility
38 Sidney Street, Suite 100
Cambridge, MA 02139
U.S.A.
IMPORTANT: Please note that we are not receiving samples on Saturdays and Sundays or on official holidays.