Laragen Primer Walking Sequencing, DNA

Primer Walking Sequencing Services

Our primer walking service includes primer design and synthesis, sequencing reactions and sequencing assembly. Our low price primer walking sequencing services only charges for the cost of the primers and the purified template sequencing reactions.

Sample Requirements:

Customers can send us 20 to 30 ul of purified plasmid and PCR DNA 1.7 ml Eppendorf tubes and 5 to 10 ul of 10 uM original primers (if any) in separate tubes. Please label on the side of the tubes. Laragen will design and order the necessary primers to finish the complete sequences. In most of cases, we can get 700 to 800 bp per primer read. To request this service, please type primer walking in the comment field when submitting the request form online.

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 clients who are not covered by free our pickup service, please ship your samples at room temperature to:

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