User:Meaganl/Online DNA submission
From 2007.igem.org
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===Online Submission Form=== | ===Online Submission Form=== | ||
| + | In order to accept physical DNA submission from teams in an organized fashion, we will be requiring teams to complete an online DNA submission form. The form is comprised of four parts: | ||
| + | ====Preliminary part specification page==== | ||
| + | |||
| + | This page is the first page you will come to when you access the online submission form. You will need to specify which one of the four formats you are sending your parts in to the Registry: | ||
| + | |||
| + | * '''Single PCR tube''' - If you are sending less than 4 samples, you may use the single PCR tube format to send in the physical DNA for your parts. The tube(s) must be wrapped with lab tape with the sequential number of the part written in permanent marker on tab of tape (this number is NOT the part number - it is the number of the sample i.e. #1, #2, #3, #4). When submitting 8-tube strips you must ship them in 50ml Falcon tubes. This format requires a total of 20ng of miniprepped DNA in a final volume of 10ul per sample. | ||
| + | |||
| + | * '''8-tube strip''' - To send more than 4 parts at a time, use the 8-tube strip format. Wrap lab tape around the first tube in the sequence then write the part sequential number (see above) on the lab tape tab. When submitting 8-tube strips you must ship them in 50ml Falcon tubes. Each Falcon tube can hold 2 8-tube strips. ''Using the 8-tube strip format requires a total of 20ng of miniprepped DNA in a final volume of 10ul per sample.'' | ||
| + | |||
| + | * '''96-well PCR plate''' - If you are submitting a large number of samples you may wish to submit the DNA in 96-well PCR plate formate. You need to add the samples in the correct order. Start with well 1A and work your way down the first column. Then continue on to well 2A and down that column. Proceed in this fashion until you have added all of the samples that you are submitting. Wrap the well 1A with lab tape to provide yourself as well as the folks at the Registry with a visible marker as to where to start processing samples. ''Using this format requires that you submit 20ng of miniprepped DNA in a final volume of 10ul per sample.'' | ||
| + | |||
| + | * '''Filter paper grid''' - We have put a lot of effort into testing the use of high quality paper as a method by which to send DNA. The Registry will be sending each team several paper grids onto which they can spot their DNA. You must combine 1.6ul of DNA at 100ng/ul with 0.4ul of 1% Cresol Red dye and spot the resulting 2ul in the middle of the box into which you are spotting your DNA. Begin with box 1A, continue down column 1, continue on to box 2A and down that column. Proceed in this order until you have finished spotting all of the samples that you are submittin. ''You will end up submitting at least 160ng total for each sample.'' | ||
| + | |||
| + | |||
| + | After choosing which method for DNA submission you will use, you will need to enter the Registry part numbers for each sample. Enter the part numbers in the order in which you will be sending the DNA. By default this page will only 8 text fields into which you can input your part numbers. If you are submitting more than 8 parts, click on the appropriate button to '''Add more parts.''' | ||
| + | |||
| + | Once you have finished entering the part numbers for all of the parts that you are submitting, click on the '''Proceed to fill in part details''' button. This will take you to the next portion of the online submission process. | ||
===Physical DNA Submission: Screening stages=== | ===Physical DNA Submission: Screening stages=== | ||
Revision as of 15:56, 23 August 2007
Contents |
iGEM 2007 Part Submission
In order to qualify for winning any part-related awards at the 2007 iGEM Jamboree, the teams must document their parts according to the Part Documentation Guidelines [PROVIDE LINK] as well as submit the physical DNA to the Registry. Following these guidelines will begin the part promotion process to get your part elevated to the Accepted rating level. Only parts that are at the Accepted rating level will be considered for awards.
For iGEM 2007 we are accepting part submissions in the form of miniprepped DNA. There is an online submission form that must be completed in order to start the DNA submission process (see more details below) [PROVIDE LINK TO FORM]. Each part will go through a physical DNA screening/quality control process, in addition to the part documentation review process, before it is accepted into the Registry. We will also provide online status updates for each part during the submission process so that the part creator can find out at what stage of the screening process their part has gone through.
Part Documentation Guidelines
Only parts that are deemed Accepted by the Registry will be considered for part-related awards in the iGEM 2007 Jamboree. In order to get your part promoted to the Accepted rating level, you must follow the instructions on the PART PROMOTION PAGE.
Quality Control Process
In order to assess the quality of the parts accepted into the Registry, we will perform a standard set of experiments with a focus on verifying the length of the insert. In addition we will aim to validate the declared antibiotic resistance of each part.
The quality control process is comprised of the following tasks:
- Transform: we will transform each sample of DNA into our standard cell strain, Top10
- Pick/Inoculate single colony: after transforming the DNA into Top10 cells, we will pick a single colony and inoculate into the antibiotic specified by the creator. We will also inoculate that colony into the rest of our standard antibiotic set (Ampicillin, Chloramphenicol, Tetracycline, Kanamycin) and watch for growth in the correct antibiotic
- Miniprep : the bacterial culture grown in the specified antibiotic will be miniprepped to isolate plasmid DNA
- Digest : the miniprepped plasmid DNA will be cut with EcoRI and PstI to cut out the insert from the plasmid
- Gel: the digested plasmid DNA will be run on an Invitrogen E-gel to visualize the insert and the plasmid. From this gel we can tell whether the part length corresponds to the length designated by the designer.
If the part length as detected on the E-gel corroborates what the designer has specified in the part documentation AND if the antibiotic testing is consistent with the expected antibiotic growth pattern, the physical DNA for the submitted part will be deemed Accepted.
Online Submission Form
In order to accept physical DNA submission from teams in an organized fashion, we will be requiring teams to complete an online DNA submission form. The form is comprised of four parts:
Preliminary part specification page
This page is the first page you will come to when you access the online submission form. You will need to specify which one of the four formats you are sending your parts in to the Registry:
- Single PCR tube - If you are sending less than 4 samples, you may use the single PCR tube format to send in the physical DNA for your parts. The tube(s) must be wrapped with lab tape with the sequential number of the part written in permanent marker on tab of tape (this number is NOT the part number - it is the number of the sample i.e. #1, #2, #3, #4). When submitting 8-tube strips you must ship them in 50ml Falcon tubes. This format requires a total of 20ng of miniprepped DNA in a final volume of 10ul per sample.
- 8-tube strip - To send more than 4 parts at a time, use the 8-tube strip format. Wrap lab tape around the first tube in the sequence then write the part sequential number (see above) on the lab tape tab. When submitting 8-tube strips you must ship them in 50ml Falcon tubes. Each Falcon tube can hold 2 8-tube strips. Using the 8-tube strip format requires a total of 20ng of miniprepped DNA in a final volume of 10ul per sample.
- 96-well PCR plate - If you are submitting a large number of samples you may wish to submit the DNA in 96-well PCR plate formate. You need to add the samples in the correct order. Start with well 1A and work your way down the first column. Then continue on to well 2A and down that column. Proceed in this fashion until you have added all of the samples that you are submitting. Wrap the well 1A with lab tape to provide yourself as well as the folks at the Registry with a visible marker as to where to start processing samples. Using this format requires that you submit 20ng of miniprepped DNA in a final volume of 10ul per sample.
- Filter paper grid - We have put a lot of effort into testing the use of high quality paper as a method by which to send DNA. The Registry will be sending each team several paper grids onto which they can spot their DNA. You must combine 1.6ul of DNA at 100ng/ul with 0.4ul of 1% Cresol Red dye and spot the resulting 2ul in the middle of the box into which you are spotting your DNA. Begin with box 1A, continue down column 1, continue on to box 2A and down that column. Proceed in this order until you have finished spotting all of the samples that you are submittin. You will end up submitting at least 160ng total for each sample.
After choosing which method for DNA submission you will use, you will need to enter the Registry part numbers for each sample. Enter the part numbers in the order in which you will be sending the DNA. By default this page will only 8 text fields into which you can input your part numbers. If you are submitting more than 8 parts, click on the appropriate button to Add more parts.
Once you have finished entering the part numbers for all of the parts that you are submitting, click on the Proceed to fill in part details button. This will take you to the next portion of the online submission process.
