Method Identifying Hybridizing Regions of DNA Within an Insert

摘要

A common problem arising during screening of DNA libraries using heterologous probes and low stringency hybridization conditions is that inserts which have only a short stretch of sequence identical to the probe can hybridize to the probe. One has torely on sequence comparisons in order to distinguish an authentic gene from false positives. However, sequencing inserts of several kilobases of DNA from 10 to 20 clones is a time-consuming task. Here we present a novel technique which can quickly identify the DNA region that causes hybridization to the probe, so that one only needs to sequence a 200-300-bp region to locate the source of the hybridization signal.