Preparation of Primer

Primers should be at least 18mer and desalted. Each sample will require 10 picomoles primer per reaction. Calculate the melting temperature of your primer. One of several methods is:

Tm = [(A+T) x 2°C] + [(G+C) x 4°C]

This will determine the cycle sequencing program used to incorporate the fluorescent dideoxynucleotides.
10 pmole/rx (1ul of 10uM) sequencing primer is used.

Optimal results are obtained when primers are diluted in dH2O 10mM Tris buffer (Avoid TE buffer)

Comments are closed.