Sequencing

From FreeBio

{The following describes the steps for preparing DNA samples for sequencing.}

GENEWIZ

  1. Fill out the sequencing submssion forms, which can be found at link.
    {Remember to keep a record of what each ID label corresponds to! I keep a spreadsheet w. lisitng the ID, the name of template DNA, the name of primer and the length of the template.}
  2. Label 8-tube PCR strip-tubes w. a 2 letter initial followed by a three digit numeral, e.g. YL001.
  3. For each sequencing rxn, mix the following:
    • 8µl template DNA
    • 4µl 2µM sequencing primer
      • Prepare 2µM primer stock by adding 1 volume 10µM primer stock to 4 volumes ddH2O.
        {Thus, to obtain 50µl 2µM primer stock, mix 10µl 10µM primer stock with 40µl ddH2O.}
  4. Close the strip-tubes w. strip-caps, and seal w. parafilm.
  5. Give to Alain.
  • Sequencing submission forms for GENEWIZ can be found at link. Sample preparation guidelines can be found at link.

Posted dna1112 18:04, 23 Jun 2005 (EDT)