Sanger DNA Sequencing

• $6.00 per sequencing reaction for standard Sanger sequencing
• Universal primers are available at no charge for use in sequencing.  Please see our list of universal primers using the “Universal Primers” link at the bottom of the page.  
• PeakTrace_­ is being used to give you longer reads of approximately 1000 bases
• The new order form (excel format) is found on the home page at the top.  Please download the form, fill it out, and then email it to us just prior to bringing your samples. 
• 
• Samples received before 1:00 p.m. are available in the afternoon/evening of the same day (please see the whiteboard as sometimes this may be subject to change due to personnel constraints)
• Primer design is available to sequence your insert or entire plasmid.  Please see the link at the bottom of the page entitled “Project Requests” 
• Sample Drop Box:
  •  Outside of Room 418 WBSB.  Samples can be left in one of the racks inside the small white refrigerator.  
• We have a variety of ways to help you save money, please inquire if you are someone who brings a lot of samples at one time.
• You can look at your chromatogram files (.ab1 files) by using Chromas.  You can find the download below.  Copy and paste the URL into your browser. 
• https://chromas.software.informer.com/2.5/

We can clean-up your PCR products and take them straight into sequencing using a robotic bead clean-up method.  Please inquire with us if you have not used this service before.

• Cost:  $1.50 per sample

• Plasmid Templates:  0.1 to 0.2 ug/ul in water – 7 ul minimum per reaction
• PCR Products:  10 to 15 ng/ul (with an average length of 500 bp) – please adjust your concentration accordingly for longer PCR products.  If your PCR products are large, i.e., 4Kb or something like that, then use the amounts requested under Plasmid Templates – 7 ul minimum per reaction.

• Do you want us to clean your PCR products for you?  Please see the PCR product clean-up section above.
• Large DNA, i.e., Genomic DNA – 0.5 ug/ul with 12ul per reaction in water.  This DNA must be very clean.  The charge for this is $23.00 per reaction.  PLEASE DO NOT MIX THE TEMPLATE AND PRIMER TOGETHER.

• Custom Primers supplied by you:  4.0 uM in water – 7 ul minimum per reaction with the exception of Large DNA runs.
• Custom Primers for Large DNA runs:  8 uM in water – 12 ul minimum per reaction.

If you have any of the following known areas in your template, please mark the order form accordingly.  Pricing is also listed below for difficult template sequencing.  For hairpin, base repeats, and shRNAi, we are using a special sequencing mix to help get through the difficult regions.

• GC-rich templates:  A high GC content in a small area of your template (65%+) (Check the GC rich box on the order form)
  • No added charge
• Hairpin Structures: A sequence containing two inverted repeats that are separated by at least three nucleotides (check the RNAi box on the order form)
  • $1.00 added charge

• Base Repeats:  Di- or tri-nucleotide repeats (check the RNAi box on the order form)
  • $1.00 added charge

• If you desire to mix your template and primer together in one tube, please use the concentrations requested.  For example, add 7 ul of the template at the required concentration and 7 ul of the primer at the required concentration into the tube and mark the “Mixed” column on the order form.  At this point you should have 14 ul in the tube.
• As a reminder, if you are doing Large DNA sequencing, please do not mix.

Plasmid Template Preparation

Retrieving Data From the Server

Universal Primers

Project Requests (sequencing long inserts)

Please feel free to contact us: Lab: (410) 955-2739 | e-mail: syn_seq@jhmi.edu