We used the BigDye kit for sequencing that comes from Applied Biosystems

It is based on a modern version of the Sanger method for sequencing, where a mix of fluorescent ddntps and dntps is used.


Usually we sequence following the recommendations from the commercial information:

VLupa.jpg we have had good results with a total volume of 8 uL

  • Sequencing
total DNA 500 ng
Mix 2 uL (this mix has: dntps, ddntps, polymerase and buffer)
primers 1uL
ddwater up to 8 uL

Fluorescence screening

we read the samples in a 96-capillar fluorescence spectrometry.