Sample Requirements for
Sanger Sequencing/San Diego

Here’s how to prepare your samples for sequencing.

Volume

We need 5µl per reaction of both your primer and DNA at the concentrations specified below:

Coming Soon: For Colony PCR (plate only service)
Plasmid DNA	100ng/µl
PCR product	10ng/µl
Sequencing Primer	3.2pmol/µl
Colony PCR Liquid Culture	150µl of overnight culture provided in a 96-well plate
Colony PCR Primers	100µl at 10pmol/µl per plate
Sequencing Primers	300µl at 3.2pmol/µl per plate

DNA Concentration Needed

Primers

We require your primers to be sent at 3.2pmol/µl and require 5µl per reaction. You can choose to send your own primers, use our stock primers, or you can order custom primers that will be delivered to the sequencing laboratory.

We recommend the following when designing your primers:

Quality of Sample

All samples must be of a good quality, free from contaminants such as salts, other contaminating DNA, unincorporated dinucleotides (in PCR reactions), and other reagents that would interfere with the reaction.

Prior to submission, we advise running your samples on agarose gels to ensure only one clearly defined band is visible on the gel (i.e. that your DNA is not contaminated with other DNA/it has not degraded).

It is important that you are aware of any motifs or secondary structures that might impede high quality sequencing results. If you know that this is most likely going to be a problem, we recommend you use our Difficult Template Protocol (dGTP) option when ordering.