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Do you have any additional drop-off locations for sequencing?
We do not have any additional drop-off locations for submitting samples. All samples must be sent to or brought to the 10th floor of the Enders Building in room 1050.
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Several members of the lab are interested in using the Molecular Genetics Core Facility for sequencing. Can we set up one single account or should each lab member create one individually?
It is better if each individual lab member creates a separate account. If the facility needs to contact users about sequencing data or samples, it is easier and better for everyone for the facility to have direct contact information to the person submitting the samples.
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How do I site the Molecular Genetics Core Facility in a paper?
When submitting a paper to a publication, if any of the services within the core facility were utilized, it is important that you acknowledge our facility, the work we performed, and the grant we receive from the Mental Retardation and Developmental Disabilities Research Center (MRDDRC). Our full name is the MRDDRC Molecular Genetics Core Facility at Children's Hospital Boston and the grant number is NIH-P30-HD18655.
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Is there any way to edit my sample submission? I just electronically submitted a new sample request and realized I did not include all of my samples.
Once a sample request has been generated and accepted it cannot be edited. If there are changes that need to be made, post a note on the incorrect sample request informing the facility of the error and then submit a new, correct form. If there are simply samples missing from the first request, create another one that contains only the missing samples.
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Between what times can samples be dropped off for sequencing?
The Molecular Genetics Core Facility is open from 9 am - 5 pm Monday through Friday. Samples can be dropped off at anytime during open hours. The facility is locked during all other times.
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What software can I use to view my chromatogram results?
There are many sequence viewing software programs on the market, and almost all of them can be used to view your results. There is a list of some very basic, free programs with links to websites for downloading under the "Helpful Links" header on our website.
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How do I sequence through a poly N in my template?
Often, large poly N sequence within a template uses up that specific di-deoxy nucleotide, resulting in noisy sequence after it. It is easiest to design primers right after the poly N sequence.
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My sequence is very strong at first, but then trails off quickly - how do I fix this?
Due to the nature of dye terminator cycle sequencing, a large amount of template DNA results in a large amount of starting points for your primers. This results in most of the fluorescently dyed nucleotides to be incorporated at the beginning of the template. In these cases, we recommend customers to submit their template DNA at a lower concentration.
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How do I sequence through a G-C rich area or a hairpin in my template?
We suggest suspending your sample in 5-10% DMSO. This should be submitted in the same tube with your template and primer. If this does not work, please contact us to discuss other possible solutions.
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What length reads can I expect?
Our sequencers can produce quality read lengths up to 1000 bases. Remember that the length of the sequence relies heavily on the quality of the template submitted.
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Do you supply any sequencing primers?
We do NOT supply any primers for sequencing.
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What type of tubes or plates do you recommend we use to submit samples?
We ask that all our customers submitting individual samples use 200ul striptubes with unattached stripcaps. These are the ONLY types of tubes that work with our current protocol. For full plate sequence submissions, we ask that customers submit samples in 96-well, flat-surfaced, skirtless plates enclosed with an adhesive film. If you need specific ordering information, feel free to contact us.
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Do I add my reverse primers and forward primers in the same tube?
No. You must have a separate reaction for each primer to sequence your template.
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Do I add the primer to my template or supply the primer separately?
Please combine the primer and template in the same tube. The template and primer will remain stable in the refrigerator for over a week.
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Where do I bring my sequence samples and sample sheets?
The Molecular Genetics Core Facility is located on the 10th floor of the Enders Research building at Children's Hospital Boston in room #1050. Immediately on the right when entering the lab is a refrigerator for samples and trays for sample sheets. Place your sequence samples in the provided lettered box corresponding to the first letter of your last name and then into the refrigerator. Be sure to clearly label your samples with tape, including your name and the date submitted. Place your corresponding sample sheet in the tray associated with the type of DNA template you are submitting for sequencing.
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How long does it take to process sample submissions?
It takes generally 2 days to process the samples. However, if we receive a very low amount of samples during a day, it can sometimes take up to 3 days because we run no less than a full plate of 96 samples at a time.
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