Chapter 8, Problem 17P

Summary Introduction

To determine: The sample of given DNA segment after sequenced by Sanger method was reacted with DNA polymerase and four given nucleotide mixtures in which small amount of $\text{ddNTP}$ is added. The resulting DNA was separated by electrophoresis and the location of fluorescent band is determined. The band pattern obtained from mixture 1 is shown. The band pattern of remaining three lanes of the gel.

Introduction:

Sanger method is used for DNA sequencing in which a small concentration of dideoxynucleoside triphosphate $\text{ddCTP}$ is added along with $\text{dCTP}$. It is also known as “chain-termination DNA sequencing”. It uses template DNA having oligonucleotide primers, four deoxynucleoside triphosphates, DNA polymerase and four $\text{ddNTP}$.