Submitting Plasmids for Sequencing
From Bridges Lab Protocols
- Go to https://client-seqcore.brcf.med.umich.edu/login.cgi?exp=1
- PI login: Bridges
- Dave must add you to the lab under his account, he will give you your username and the password needed
- Typically we will select Sanger Sequencing (the only low throughput option)
- Select 'Submit samples/plates'
- Select the appropriate short-code from the drop down menu at the top of this page
- Generally choose 'One-at-a-time Sample Submission' unless you have a lot
- From here enter the following for each section:
- The sample name (the sample name that mean something to you regarding the sequence)
- No discounts
- Mini-Prep Plasmid
- Whatever vector you used for this plasmid
- Size of the insert you are sequencing
- Custom Primers
- Do not save
- Do not rush
- Save and repeat with your other samples
- Once you have finished adding your samples ID codes for each sample will be generated and the last four digits of this ID are what you need to use for the tubes you are submitting, not the sample names you made
- Samples should be submitted in 1.5mL tubes
- Make sure to write these IDs very clearly
- A submission ID will also be generated and you need to know this when you drop your samples off so they can find your sequence request in their system
- Plasmids need to be diluted to a concentration of ~50ng/ul and total volume should be 10ul and should be labeled with the generated sample ID
- In addition to your samples you will need to provide an aliquot of the primer
- Volume is 10ul/sample you submit
- Should be labeled as the range of sample IDs generated and P1 (example: xxxx-xxxxP1), if you have more than one primer write xxxx-xxxxP2 and so on
- Go to https://seqcore.brcf.med.umich.edu/sample-dropoff-locations for locations and drop off times
- They will send you an email when the project has been completed. You can download the .ab1 files here: https://client-seqcore.brcf.med.umich.edu/ftpdata.cgi
