Sanger Sequencing FAQs

  1. How should I provide my template and primer? Please provide your pre-mixed sample containing 6 µl of template and 6 µl of primer, at the following concentrations:
    1. Primer: 1 µM for PCR or Plasmid, 4 µM for BAC.
    2. PCR products: 3.0 ng/µl/100 bp.
    3. Plasmid DNA: 0.2 µg/µl.
    4. BAC/PAC DNA: 0.5 µg/µl.
  2. How much template should I bring if I want to use your primers? If using the primers available at DAF, please bring 6µl of template in a tube and add 6µl of the appropriate primer when you arrive at our facility. There is no charge for using our primers.
  3. How do I place a sequencing order? The JHU ordering server manages all orders and data. To place an order and view your data, you must have a username and password. If you do not have an account, you can set one up from the log-in page. Please use your PI’s name instead of the Institution to help us with the final step of the set up.
  4. Should each person in the lab have their own log-in?  In order to utilize all of the features of the server, including order notifications and data tracking, each individual in your lab should have their own login.  If your lab does not wish to do this, keep in mind that we will contact the person whose name is on the account with any questions or concerns.  This may cause a delay in processing your samples.
  5. When will my data be ready?Results for sequencing samples dropped off in Blalock by 4:00 PM will be available by 10:00 AM the following morning, assuming no machine problems. Samples dropped off in our remote locations by 11:00 AM will receive their data by 10:00 AM the following morning. You will receive an email when your data is completed.
  6. Where do I drop off my samples? 
    1. Blalock 1005: Open 7 AM – 4:30 PM. There is also an after hours drop-box available for after-hours.
    2. CRB I B02A (slot A63): Samples should be dropped off by 11AM, though on most days samples dropped off here by 2:00 PM will picked up and processed the same day, making the data available the following morning.  First time users will need to email Lee Hilliard or call 410-502-3959 to set up and account. You may also download forms for 24/7 access. If you already have a 24/7 account, you can just login to the system and drop off the sample. Someone is usually available at the 24/7 from 3:00 PM to 4 PM to help with problems and questions.
    3. Bayview Campus: Drop off in the Core 24/7 on the first floor of the Asthma and Allergy building, in slot F51. Pickup is once daily. Samples dropped off by 11:00 AM will have data available by 10 AM the next morning. Samples dropped off after 11:00 will wait for the next day. First time users will need to email Lee Hilliard or call 410-502-3959 to set up and account. You may also download forms for 24/7 access. If you already have a 24/7 account, you can just login to the system and drop off the sample. Someone is usually at the 24/7 from 11:00 AM to 12 PM to help with problems and questions.
  7. How should I submit my samples?
    1. Tubes: due to storage issues, we will accept only 1.5 ml eppendorf tubes. Plates: 24 samples or more may be submitted on plates with the requirement that all consecutive wells are filled in a top to bottom, left to right format (i.e. A1-H1; A2-H2, etc.).
    2. Labeling: Your tubes need to be labeled with your Genesifter Tracking ID and the position number.
    3. Note: Any sample that is improperly labeled, unreadable, or any sample sets organized in a way that we feel may increase the risk of error during handling may be rejected. Improperly labeled samples will be returned.