Purified Template Sequencing Service
By far, this is the most common service we provide. With this service, you provide us purified plasmid or PCR DNA templates with or without DNA concentration. Unlike most of our competitors, which depend on customer's DNA concentration, we measure DNA concentration of every single sample with our fluorometer. Then we will adjust the amount of DNA used per reaction based on the DNA concentration we measure. This procedure enables us to have good sequencing quality and long reads.
Customers can send us 10 to 15 ul of purified plasmid and PCR DNA either in 8 strip PCR tubes or in individual 1.7 ml Eppendorf tubes and 5 to 10 ul of 10 uM primer in separate tubes. Laragen provides free common sequencing primers. Please do not label the PCR strips on the caps. Please label on the side of the tubes.
The turnaround time is overnight. Average reads can be up to 1000 bp depending on DNA quality and DNA and primer sequences. We will provide a free repeat for failed reactions for this service.
The key to having long sequencing reads is the quality and quantity of DNA. Too much or too little DNA and dirty DNA are the main causes of the reaction failure. For PCR DNA, there should be a single band on an agarose gel. We need 10 ng of PCR DNA for every 100 bp of PCR products. Please check PCR DNA concentration after purification. Many commercial PCR kits are not robust and the recovery yield varies from sample to sample. For plasmid DNA, the optimal DNA concentration is ~100 ng/ul. We recommend Qiagen miniprep kit to prepare plasmid DNA. Certain commercial kits did not work well with automated Sanger sequencing. EDTA will inhibit DNA sequencing reaction. Don't elute DNA into TE buffer. Use only water or Qiagen EB buffer (10 mM TrisHCl, pH 8.0).
For 96-well plate, please arrange samples by the columns, not by the rows. You can use upload and import file function in our DNALIMS system to upload 96-well plate format. No spaces or illegal characters allowed in the DNALIMS. Allowable characters include: a - z, A - Z, 0 - 9, and hyphens. You can use this template for file upload.
Please seal the 96-well plate with caps, foil tape or heat seal tape to avoid possible cross contamination and sample evaporation. Please label on the side of plate. Don't label on the top (seal or caps) of the plates.
If you aren't using our convenient free pickup service, ship your samples at room temperature to:
10601 Virginia Ave.
Culver City, CA 90232