sample submission guidelines

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When filling out the form, please keep the following in mind:

• Sample names cannot have spaces, but may include the following "special" characters:  -+.(){}_#
• Sample names must be 35 characters or less. We prefer your initials and a number.
• If submitting plates, please provide a unique plate name or barcode on the order form, and write it on the side of the plate!

DO:

• WRITE CLEARLY
• USE SHORT, DISTINCT NAMES FOR ALL SAMPLES
• USE 1.5 mL TUBES (PICTURED ON LEFT). 0.6 mL tubes are also acceptable, but we prefer the larger tubes if at all possible. 
• IF SUBMITTING MORE THAN 32-48 SAMPLES, PLEASE CONSIDER SENDING US A PLATE AS OPPOSED TO TUBES.  
• ADD YOUR OWN PRIMER UNLESS WE ARE NEEDING TO DO A CLEAN-UP FIRST, OR IF YOU WANT US TO ADD ONE OF OUR STOCK (FREE) PRIMERS
• INDICATE IF YOU WOULD LIKE (OR NEED) THE dGTP PROTOCOL (SEE BELOW)
• INCLUDE THE PRINTED FORM ALONG WITH YOUR SAMPLES
• INCLUDE CUSTOM PRIMERS WITH THEIR CONCENTRATION AND CLEAR LABELING
• USE BUBBLE WRAP OR STRONG SECONDARY CONTAINERS FOR SHIPPING (50mL TUBES, ETC). 
• USE UPDATED FORMS IF POSSIBLE (FORMS AVAILABLE HERE)
• REACH OUT TO US IF YOU HAVE ANY QUESTIONS BEFORE SEQUENCING

DO NOT:

• USE PARAFILM
• FORGET THE ORDERING FORM
• FORGET TO UPDATE YOUR INFORMATION AT THE TOP OF THE ORDER FORM(S)
• SEND SAMPLES ON DRY ICE (THEY CAN IN FACT BE SHIPPED AT ROOM TEMPERATURE)
• WRITE SO SMALL THAT WE CANNOT READ IT

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:

• ssDNA plasmid:  0.25μg DNA/5kb    + 2 pmol primer in 13 µl
• dsDNA plasmid:  0.5μg DNA/5kb      + 8 pmol primer in 13 µl
• PCR products:    2-4ng DNA/100bp  + 8 pmol primer in 13 µl

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: 

• 1µL of 10µM primer, or 
• 3.2µL of 3µM primer 

If requesting one of our stock primers, please CLEARLY INDICATE for us to add the appropriate primer in the special instructions column of our order form, and submit 10uL total volume of your DNA samples (without primer).

Template: Please be sure that you do not have SALTS, PROTEIN, DETERGENTS, RNA, CHROMOSOMAL DNA, ETHANOL, or EDTA. Pure water is best!

Quantify your products on an agarose gel, a NanoDrop, or a Qubit. Please let us know  if your products are at a low concentration and/or are larger than 10kb. We can quantify your DNA for an additional charge. 

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 for our purposes if at all possible, and generally works well for most DNA samples.

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 (or H12) empty for our positive control.

We require samples to be submitted in a specific type of plate:

• Catalog#: 4ti-0770/c
• Description: FrameStar 96, semi-skirted
• You may purchase these plates at the DNA Sequencing Facility for $6 each
• If you are shipping plates to our Facility, please use very tight strip caps.

When submitting samples in a plate, the total volume per well should be 4µL that includes:

dsDNA plasmid: 250ng/5kb  + 4 pmol primer or PCR product: 20ng/1000 bp  + 4 pmol primer. Seal the plate well, and bring it to us. We will spin it down and proceed with sequencing... and will let you know if anything looks awry. 

Please download the Full Plate Sequencing Order Form, e-mail us a copy, and print a copy to include with your plate. 

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 and/or in the Special Instructions field.

We have changed our standard protocol to provide the best possible results... including for GC-rich templates. We no longer require customers to indicate "dGTP" for processing, as we do that automatically. That said -- if you KNOW that your sample has meaningful secondary structure / stretches of high amounts of G's and C's, please do let us know. 

Please refer to our Nanopore sequencing service page.