Sanger DNA Sequencing

Overview:

Instrumentation consists of an Applied Biosystems 3730xl DNA Analyzer.

NEWS: 

  • DNA Sequencing $6.00/reaction
  • PeakTrace is now being used to give you longer reads of around 1020 good bases. 
  • We are offering Ampure PCR clean-up using our robot for $1.50 a sample.

If you drop your sample off by 1:00 p.m., you can usually expect your data to be ready the next day.  If we do not have a full plate at 1:00, we usually will process the samples the next day in the morning and in most cases the data will be out that afternoon.  Please look at the white board when you drop off your samples for the date your data will be ready.  The date listed is the worse case scenario, it is always a good idea to check the server the next day in the afternoon anyway no matter what time you drop your samples off. 

We also offer Saturday delivery of your data to the server.

Sequencing Cost:  $6.00 per reaction (includes universal primer -- see universal primer link below for a complete list of our free primers.  We can add to the list if there is a primer that you think we should stock.)

Note:  For samples with an RNAi insert, we charge $1.00 more per sample.  Large templates such as BACS are $23.00 per sample.  

Campus Sample Drop-Off Locations:

We have three drop-off locations located on campus. 

WBSB:  Outside of Room 418 for after hours.

BRB:  Outside of room 372

SOPH:  Inside the Monument street entrance where the FedEx box is located.  Look for the black "mailbox" on the wall.  Instructions are on the box.

Requested Concentrations:

Custom Primers Supplied by You:   4.0 uM in water with a total of 7 ul per reaction

Plasmid Templates:   0.1 to 0.2 ug/ul for (ds) plasmids with 7 ul per reaction in water.

ShRNAi Templates:  0.2 ug/ul with 10 ul per reaction in water.  DO NOT MIX WITH PRIMER, please provide template and primer separately if using custom primers.

PCR Products:  10 to 15 ng/ul (with an average length of 500 bp) adjust amount accordingly for longer PCR products.  If your PCR products are large, i.e., 4 Kb or something like that, then use the amounts requested under Plasmid Template.

BACs and COSMIDs:  0.5 ug/ul  with 12 ul per reaction in water.  Must be very clean.  The charge for this is $20.00 per reaction.  DO NOT MIX WITH PRIMER.

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The benefits of using YOUR JHMI Sequencing Facility:

  • First and foremost, we are here for YOU.  This is the sole reason the facility exists.  So please make use of your Hopkins facility. 
  • Template and Primer can be mixed by you or we can do it here in the lab. There is no extra charge if we mix it.  If mixing, use equal amounts. 
  • We also offer other discounted pricing for samples that are already cycle sequenced by you.  Ask the staff for more details. 
  • All samples will be stored for a month.  We will store your samples longer if you request this.  We also will hold on to primers that you use frequently as well. 
  • Every chromatogram is analyzed by the staff.  We will repeat samples, at our discretion, should we see that something failed when everything else worked. 
  • We are available to talk with you from 7:30 a.m.  to 5:00 p.m. Monday through Friday regarding your sequencing. 


    Free DNA Sequencing Chromatogram Viewers:

   Applied Biosystems

       Sequence Scanner

For more detailed information please select one of the following:

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 | Fax: (410) 614-7566 | e-mail: syn_seq@jhmi.edu

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