Your DNA template must be very clean for successful sequencing. Make sure you follow recommended methods of preparation and adjust your concentrations properly. Don’t assume that because a particular kit or protocol for isolation works for another lab procedure, it will work with our system. It may not. See our Sample Preparation page for information on preparing template.
Resuspend your DNA template in very clean dH2O. The tris buffer that comes with the Qiagen kits will work if it does not contain EDTA. Do not use TE! The EDTA cheletes the magnesium ions required for our enzyme to work.
Do not use JM109 as a host strain. It makes excessive amounts of protein and it is difficult to get a preparation free of protein contamination.
Protein contamination and improper concentrations are the most frequent causes of inadequate sequence. In addition, improperly prepared sample can be detrimental to our capillary array. Please make every effort to give us high quality samples!
Qiagen publishes a very good template purification guide. It would serve you well to download and read it. Please ignore its advice on genomic preparation as we do not accept genomic preps.
Concentrations and Volumes
Concentrations are important! Too much will shorten your sequence and too little will Please don’t submit samples that are not at the requested concentrations!
Plasmids: 50-75 ng/ul and 4-5ul for every reaction.
PCR Products: Please see our Sequencing PCR Products Page for concentrations and volumes.
Where to send your samples:
All requests must be submitted through iLab. See the links below for your particular institution.
Please provide a gel documentation image of your samples, concentration, and 260/280 ratios. This is for QC purposes and we will not sequence your samples without this information.
Bring or ship your samples to: