I’m also sequencing PCR-Products using Alf Express.
I clean the PCR-Product from the old primers using the “PCR-kit” from GE.
This product is used directly without any ss-ends.
Instead of cy5-dATP I have sequencing primers 5’labeled with cy5.
The sequencing primer are positioned like a “nested PCR-primer” a little bit
away from the end of the PCR product.
The sequencing reaction is carried out using the “Sequencing-Kit” from GE.
If you are interested in the protokoll, please let me know.
Saluti and ciao
Sabine Lautenschläger