Valencia/Sequencing
From 2007.igem.org
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.
Sequencing
Usually we sequence following the recommendations from the commercial information:
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.