Laragen Sequencing Genotyping

Difficult Template Sequencing Services

Using Laragen's proprietary protocols and techniques, we can handle your difficult templates! It will expedite the sequencing results if the you inform us if the sequence contains the following structures: shRNA, RNAi, hairpin, GC rich motif, or AT rich motif. Please check GC field in the online request form when submitting these sequences. Additional charges may apply to these templates.

The turnaround time is overnight. Average reads can be up to 800 bp depending on the DNA quality and DNA primer sequences. We will provide a free repeat for failed reactions for this service.

Sample Requirements:
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 along with 5 to 10 ul of 10 uM primer in separate tubes. Please don’t label the PCR strips on the caps. Please label on the side of the tubes.

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 kits 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:

Laragen, Inc.
10601 Virginia Ave.
Culver City, CA 90232