Please follow these guidelines when preparing your samples:
Each sample tube (1.5ml tubes preferred) should contain one primer and one template at the quantities specified below:
Please note that the values listed for the primer are quantities, not concentrations.
A simple and approximate usage is to include (into the total 13µL volume), either:
If requesting one of our stock primers, put a large note in red on your order form, and submit samples in 10uL total volume.
Template: Please be sure that you do not have SALTS, PROTEIN, DETERGENTS, RNA, CHROMOSOMAL DNA, ETHANOL, or EDTA.
Quantify your products on an agarose gel or use a NanoDrop. Please let us know if your products are at a low concentration and/or are larger than 10kb.
Primers: Our default annealing temperature is 50°C. Please let us know if this is not compatible with your primer(s), and please indicate the required temperature. 50°C is preferable if possible.
Order forms can be filled out and printed from this web site by following the DNA Sequencing Order Form Download Link
You may also submit samples in a 96-well plate. We charge per sample, so you don't pay for empty wells. Please keep well A1 empty for our positive control.
We prefer samples to be submitted in a specific type of plate:
When submitting samples in a plate, the total volume per well should be 6µL that includes:
dsDNA plasmid: 250ng/5kb + 4 pmol primer or PCR product: 20ng/1000 bp + 4 pmol primer.
If you are requesting the dGTP protocol due to the presumptive presence of secondary structure in your target sequence, please reduce the volume of your template+primer to 4µL.
Please download the Full Plate Sequencing Order Form, e-mail us a copy and print a copy to include with your plate.
When filling out the form, please keep the following in mind:
For templates larger than 10KB, please provide 1µg of template with 10pmol of primer in total volume of 13µL.
In addition, please write "Large Template Protocol" in large print across the order form.
IMPORTANTLY -- If you suspect that there may be a secondary strucure in your template, please indicate in BIG RED MARKER that you want "dGTP PROTOCOL"
dGTP protocol may also work with GC and GT-rich templates.
We are testing a new sequencing protocol for cosmid, BACs, PACs, and Fosmids. We will sequence your sample for free of charge. If you have a protocol you want us try, we will be happy to try it.
Please provide:
Things to consider when preparing Cosmid, BACs, PACs and Fosmid for sequencing:
sequencing_handbook_FLR (pdf)
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