AAV Genome Sequencing: Sample Submission
To ensure the successful sequencing and analysis of your rAAV 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 note! We only accept non-infectious material that can be handled at BSL1.
Please submit purified rAAV capsids only! Other Viral Genome DNA must be submitted to our Viral Genome Sequencing service, as our rAAV workflow and analysis pipeline are specifically compatible only with rAAV samples. At present, our analysis pipeline can only process single-stranded rAAV. Please contact us if you require self-complementary rAAV sequencing!
Our workflow requires the submission of 100 μl of a minimum of 3.0 x 10^11 GC of purified rAAV capsids. Please send samples in Styrofoam containers with cold packs or on dry ice.
Samples should be submitted in common rAAV formulation buffers, PBS or TBS, and quantified via qPCR or ddPCR.
Please submit the following reference sequences in FASTA format for each sample: transgene plasmid, helper plasmid, and rep-cap plasmid. Package all FASTA files into a single compressed folder (e.g., .zip or .tar.gz), and name the folder using the orderer's last name and first initial (e.g. SmithJ).
These references are required as input for our analysis pipeline. Please make sure that the FASTA file names are IDENTICAL to the file names listed in the sample submission form.Also provide the NCBI GenBank or RefSeq number of the host genome/host expression system used in producing the rAAV vectors. We will download the host genome/host expression directly to our servers as the FASTA file sizes will exceed upload limits.
Currently, we accept samples in 1.5 ml tubes or 0.5-3 ml screw top cryovials.
Tube Orders:
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If you are submitting your samples in tube format, please use standard 1.5 ml flip-cap Eppendorf-type tubes or 0.5-3 ml screw top cryovials.
Please label the top of the lid and side of each tube - as clearly as possible - with the tube id (a maximum of 5 alpha-numeric characters; e.g., 5B001). We strongly recommend to hand-write each label using a permanent marker. Self-adherent stickers might fall off during shipment, thereby making sample identification impossible.
We recommend wrapping tubes with Parafilm to prevent samples from drying.
Tubes should be submitted in order as listed on the submission form (not randomly placed, and not loose/unracked) in a rack/box designed to hold tubes. Tubes should be placed in a Styrofoam box with a cold pack or on dry ice.
Sample Shipment:
PLEASE DO NOT SEND SAMPLES AT ROOM TEMPERATURE.
Submit your samples, in a Styrofoam box with a cold pack or on dry ice to one of our remote dropboxes ( please see here) or by delivering it personally to our core facility. Please include a hard copy of the completed order form.
If your Styrofoam box is too large to fit inside the sample drop box, please label the box clearly with "Delivery to CCIB DNA Core, 38 Sidney St." so the delivery service knows to include it in their pickup.
If remote or direct sample drop-off should not be an option for you, please ship your samples in a Styrofoam box with cold pack or on dry ice, 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.
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 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.