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DNA Submission
From 2010.igem.org
From http://partsregistry.org/DNA_Submission_Instructions.
Click [http://partsregistry.org/cgi/dna_transfer/index.cgi here] to access the Online DNA Submission form.
Contents |
Sample Details
The Registry only accepts parts in the standard DNA-shipping plasmid backbone: pSB1C3. The Spring 2010 DNA Distribution kits contained samples of this plasmid backbone in linearized form for easy use. You can also make your own linearized plasmid backbone using [http://partsregistry.org/Help:Protocols/Linearized_Plasmid_Backbones these instructions].
New DNA Submission
When you come to the DNA Submission main page, you will be able to “start a new DNA submission” or “see your DNA submissions.” To begin the submission process click on “start a new DNA submission.”
You must tell us in what format you are sending your DNA. Choose a format from the menu and indicate how many samples you are sending. Click [http://partsregistry.org/DNA_Submission_Formats here] for more information about the accepted formats. You should also let us know if there are any special instructions that we must know about.
Keep in mind that there are specific DNA amount requirements that you must follow.
For single PCR tube:
- 20ng in 10ul volume (2ng/ul) minimum needed
- Wrap tube in lab tape and label with tube number
- Cap tightly and ship in 50ml Falcon tube
For 8-tube strips:
- 20ng in 10ul volume (2ng/ul) minimum needed
- Cap tightly
- Wrap 1st tube on each 8-tube strip with lab tape and write the correct tube number (e.g. # 1, 9, 17 etc.) with a permanent marker on the tape
- Ship 8-tube strips inside a 50ml Falcon tube (2 8-tube strips per 50ml Falcon tube)
For 96-well plate:
- 20ng in 10ul volume (2ng/ul) minimum needed
- Add DNA to wells in the order of down row 1, down row 2, etc. (i.e. #1 = 1A, #2 = 1B, #3 = 1C, #4 = 1D, #5 = 1E, #6 = 1F, #7 = 1G, #8 = 1H, #9 = 2A, #10 = 2B, etc.)
- Cover tightly and carefully with aluminum foil adhesive cover (e.g. Axygen PCR-AS-200).
For filter paper grid:
- Spot at least 160ng DNA onto middle of each square, going down each row in order from left to right (follow the instructions printed on the grid).
- Fill out the online submission form as well as the paper submission form (with the grid and instructions on how to send the DNA in using this format)
- Remember to carefully fold the filter paper grid according to the instructions provided on the filter paper.
Proceed to entering sample information by clicking on the button at the bottom of the page. 350px
Sample Information
At the top of the sample information page you will see your Shipment #. You will need to remember this number, in particular if you are sending your DNA in filter paper grid format. In this case the shipment # needs to be written clearly on the filter paper grid form.
Begin to enter the information for your first part, beginning with the part number. You must enter the plasmid that the part is in. Note that the software will only recognize plasmids that are documented in the Registry so if you haven’t added your plasmid to the Registry you must do so before submitting your parts. See the “Notes” section below for more details on plasmid information.
Once you have entered the part number and plasmid for your part, you can tab over to the antibiotic resistance field. The software will automatically fill in the antibiotic resistance based on the plasmid that you have entered. Please check to make sure that the resistance is correct. You may change it manually if it is not correct. The antibiotic abbreviation is the first letter of the antibiotic.
Please check any of the boxes that apply to your part if:
1. It is toxic to Top10 cells,
2. You are only sending a plasmid,
3. It is not a part or plasmid
4. Your team has sequenced this part
We need to know specifically whether your part is toxic to Top10 cell strain because it is our standard strain. We will be transforming all parts into Top10 and we need to know in advance whether this will be a problem with any of your parts. If you are sending us a plasmid, you need to check the appropriate box (see below for more plasmid information).
Please let us know by checking the box if you or your team has sequenced this part. This is not a box to have your part sequenced. At the moment we will not be collecting any sequence data or trace files but at some point in the future we will enable this feature. Please save all of your sequencing data in the event that you would like to add that information at a later date.
At this point you may add any notes that you may want to communicate with the Registry regarding your part in the comments field.
Once you have finished adding all of the information for your part, click on the “save” link and then “add another sample” button and proceed to filling out all the information as above for each part that you are sending to the Registry. The information for each sample is automatically saved to this shipment number once you click on the “save” link.
Shipment information
When you are done filling in the information for all of your samples, click on the “I am finished specifying the samples” button. 300px Be aware that your shipment has not yet been finalized (even though it has been saved). To do so, you must fill in all of your shipping information. Please provide ALL information.
If you need to obtain a tracking number for your package before finalizing your shipment, you may do so at anytime and come back to finalize your shipment once you have a tracking number. You must provide the shipping details before your information gets sent to the Registry so make sure to come back and complete the shipping details portion of the form.
When you are finished providing shipping details, click on the “I am finished filling in this information” button to send your information to the Registry.100px
This finalizes your shipment information. You will see a confirmation page that congratulates you on submitting your information. It will also show all parts that are part of that shipment number. We recommend printing out this page for your records.
At this point you can go back to the DNA Submission Registry page and click on “see your DNA submissions” to check on the status of your shipment. You will be able to see when we have received your shipment and once we have finished testing your parts, you will be able to see the results from our quality control tests.
If for any reason we reject your part we will notify you by email (see below for possible reasons your part might be rejected). The email will be sent to the user who filled out the online submission form. The rest of the parts in your shipment that passed the quality control tests will be marked as Accepted but you will have to resubmit the parts that did not pass successfully. These parts must be resubmitted as a new shipment using the online DNA submission form once again.
Reasons for failing quality control:
- Failed to transform
- Did not grow in culture
- Growth in wrong antibiotic broth in antibiotic test
- Wrong fragment size when cut with double restriction enzymes
Shipping Instructions
Please send your samples via a shipping company that will let you track the shipment. It is important to include your shipment tracking number in your submission information details. Physical DNA for parts is due before the Jamboree (see the iGEM requirements) and if we do not receive your shipment, we will track your package and give that information to the judges. If your shipment does not have tracking information and has not arrived at the Registry we will count your physical DNA as not available. You will not be able to receive a medal, as per the iGEM requirements.
*Important*: When shipping your samples, make sure to include a detailed description of the package so that it gets through customs. Write the following:
Dry/liquid DNA, non-hazardous, non-infectious, non-regulated. For research use only.
Notes
- Accepting bacteria
- The Registry does not accept bacteria shipments at this time. Please only send isolated plasmid DNA. You may contact us if you have any questions regarding this policy.
- Editing your shipment after it has been finalized
- You may edit your shipment at any point up until the time you finalize your shipment after entering your shipping details. After you have finalized your shipment and sent all parts and information to the Registry, you will not be able to edit your online submission form. If you need to make a change to your submission form information, you must contact Meagan Lizarazo at meagan [at] igem [dot] org. Please include your shipment number, your name, your team name, and the sample information that you would like to change.
- Part and plasmid specification details
- In specifying your parts you must clearly understand the relationship between parts and plasmids. You must be explicitly aware that a part does NOT include the flanking restriction enzyme sites (called BioBrick™ prefix and suffix if you are using the BioBrick™ standard for which the restriction enzymes are: EcoRI, XbaI, SpeI, PstI). These sites must instead be specified in the plasmid that your part is in. Your part, then, begins with the first letter (A, C, T, G) following the prefix cloning site of the plasmid and ends with the last letter preceding the suffix cloning site of the plasmid.
- One of the reasons the Registry needs to know exactly what the details are for the plasmid your part is in is so that we can use the correct set of enzymes for our digest portion of the quality control process. We need to be able to calculate the length of your part when cut with that particular set of restriction enzymes (especially if is a set of restriction enzymes different than those of the BioBrick™ standard. The plasmid information must also have the antibiotic resistance markers that are part of the plasmid. This information is also extremely important for the quality control testing.
- Take a look at [http://partsregistry.org/Help:An_Introduction_to_BioBricks this help page] for more information on Biobrick™ parts and plasmids and how they are defined.
Useful links:
- iGEM 2008 Part Submission Overview
- Introduction to Biobricks™: including details on part and plasmid definitions.
Have any questions? Contact Meagan at meagan [AT] igem [DOT] org.
