Sequencing products of the polymerase chain reaction directly, without purification

Abstract

An improvement over current protocols for sequencing products of the polymerase chain reaction is described. This method allows sequencing products of the reaction without performing costly, time-consuming purification steps which often result in unacceptable loss of product. Conservation of small amounts of polymerase chain reaction products which can be obtained from limited DNA sources, such as tissue biopsies, is achieved. Clarity of autoradiograms obtained utilizing this adaptation is comparable to that obtained with the original method. In addition to streamlining the amplification-sequencing procedure, this procedure can potentially be subjected to total automation.