Sanger Sequencing

• Standard sample: $7.00

• BAC protocol reactions (used on BACs, other large templates or very GC rich templates): $27

• Same day service: $32 per sample, minimum of 5 samples

• Standard primers will be added by our staff. Please indicate which primers are needed on the order form.

Samples should be provided pre-mixed with primer in 1.5ml tubes (please do not send PCR tubes). In each tube, mix 6 ul of primer and 6 ul of template. Use only one primer per tube. To achieve the best sequencing results, start with the following concentrations prior to mixing:

• Primer: 1 uM for plasmids and PCR products, 4 uM for BAC/PAC/Cosmid

• Purified PCR Products: 3.0ng/ul/100bp

• Plasmid DNA: 0.2 ug/ul, with a 260/280 ratio between 1.75 and 1.85. 260/280 ratios above 1.9 or below 1.7 will not work.

• BAC/PAC/Cosmid DNA: 0.5 ug/ul

• If using the primers available at DAF, please bring 6 ul of template in a tube and indicate which primer should be added. Please clearly indicate which primers need to be added to the tubes. There is no charge for using our primers.

The customer is responsible for quantitating DNA and mixing with primer, if using their own primers. Custom primers should be premixed with the sample prior to being dropped off. If you are using a standard sequencing primer, please indicate the appropriate primer on the order form and we will add the primer when we receive your samples.

Tube labeling

Short Separator

Tubes should be labeled with the order tracking number and the number indicating the sample’s position in the order, i.e. if it is the first sample in the list, place the tracking id and the number 1 on the lid of the tube.

If submitting 24 or more samples, you may put them into a 96 well plate PCR plate (in column order) and label the plate with the tracking ID.

The service provides standard sequencing primers for several vectors. Custom primers should be provided by the user and premixed with the template, when needed. Primers can be ordered through our facility using  one of our portals as well.

The following primers are available free of charge.

Standard Sequencing Primers

PCR Support is available for customers who don’t have the resources to amplify samples in their own lab.  Pricing is dependent on sample number and project complexity.   Please inquire.

Insert walking and other types of specialized sequencing support are also available.   Please inquire.

• Orders must be placed in Genesifter prior to drop-off.
  • Please make sure the number of tubes you drop off matches the number of tubes in your order.
  • If you need to change your order after you submit it, call us at 410-955-2836 or email us immediately.
  • Failure to do so will result in processing delays.

Sample drop off times are as follows:

• Blalock 1004: 4 PM for daily processing.
  • Samples may be dropped off after-hours in the fridge outside our door.
  • For same day results, samples must be in Blalock 1004 by 9 AM. Additional charges and sample minimums apply.

• WBSB third floor, near the Rangos bridge: place in dropbox before noon.

• Bayview Asthma & Allergy building 1st floor hallway, in the designated mini-fridge
  • Place in fridge before 11 AM.
  • Samples may also be given to CoreStore personnel.

• Homewood: Two locations. Call 410-955-2836 for the combination to unlock the refrigerators.
  • UTL room 384
    • Black refrigerator in the break room right across from the main staircase.
  • Croft Hall outside room 141
    • Black refrigerator in the hallway
  • Samples should be in either refrigerator by 11 AM

• CRB1 Rm B02A, in the specifically designated mini-fridge. 
  • Place in fridge before 11 AM.

Turnaround Time:

• Sanger samples that are dropped off in our remote locations before the daily pickup time Monday-Thursday will have data available around 10 AM the day after they are dropped off, barring any unforeseen circumstances which prevent routine pickup. 

• Sanger samples that arrive in Blalock by 4 PM Monday-Thursday will have data available by 10 AM the following business day. 

• Samples dropped off in Blalock by 12:30 PM on Fridays will have data available on Saturday.

• Sanger samples dropped off at our remote locations anytime on Friday or in Blalock 1004 after 12:30 PM will go into our regular queue and will be released by 1 PM on Monday.

• Same day sequencing should be dropped off in Blalock 1004 by 9 AM. (There is an additional charge for same day sequencing.)

• Failing samples will be rerun on the following business day.

The  JHU ordering server (Genesifter) automatically processes data after electrophoresis is complete and provides quality assessment for each sample. Data is available for download, viewing, and editing.  Please note that Genesifter is only accessible within the Hopkins environment.  To access Genesifter from a remote location outside the JHMI domain firewall, log in to your JHED account and follow the instructions in the sidebar VPN link. Once connected through Pulse Secure you will be able to access Genesifter using the link below.