Figure 27.1: Principle of Di-Deoxy Method.

Step 5: A chase of polymerization reaction is performed in the presence of high concentration of NTPs to extend all non-terminated sequences into high molecular weight DNA. These high molecular sequences will not enter into the sequencing gel.

The different steps in labeling/termination protocol differ from sanger protocol after step-1 and it has following steps:

Step 2: A limited amount of NTPs are added along with the one of the radiolabeled nucleotide to label the DNA through the length.