I do a pre sequencing purification using EXOSAP-IT and I sequence using the Big Dye terminator cycle sequencing kit v.3.1. I then clean up using Sephadex G50 on a multi-screen filter plate before sequencing.
Please find the answer in attached-pdf on page 5.
http://dnasequencingcore.ucdenver.edu/pdf-Files/DNA%20Sequencing%20Troubleshooting%20Guide.pdf
Thanks Deden, I'll check on the document attached.
Thank you Pia. I was able to optimize my reactions finally.
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