<P> After a genomic library is constructed with a viral vector, such as lambda phage, the titer of the library can be determined . Calculating the titer allows researchers to approximate how many infectious viral particles were successfully created in the library . To do this, dilutions of the library are used to transfect cultures of E. coli of known concentrations . The cultures are then plated on agar plates and incubated overnight . The number of viral plaques are counted and can be used to calculate the total number of infectious viral particles in the library . Most viral vectors also carry a marker that allows clones containing an insert to be distinguished from those that do not have an insert . This allows researchers to also determine the percentage of infectious viral particles actually carrying a fragment of the library . </P> <P> A similar method can be used to titer genomic libraries made with non-viral vectors, such as plasmids and BACs . A test ligation of the library can be used to transform E. coli . The transformation is then spread on agar plates and incubated overnight . The titer of the transformation is determined by counting the number of colonies present on the plates . These vectors generally have a selectable marker allowing the differentiation of clones containing an insert from those that do not . By doing this test, researchers can also determine the efficiency of the ligation and make adjustments as needed to ensure they get the desired number of clones for the library . </P> <P> In order to isolate clones that contain regions of interest from a library, the library must first be screened . One method of screening is hybridization . Each transformed host cell of a library will contain only one vector with one insert of DNA . The whole library can be plated onto a filter over media . The filter and colonies are prepared for hybridization and then labeled with a probe . The target DNA - insert of interest - can be identified by detection such as autoradiography because of the hybridization with the probe as seen below . </P> <P> Another method of screening is with polymerase chain reaction (PCR). Some libraries are stored as pools of clones and screening by PCR is an efficient way to identify pools containing specific clones . </P>

Application of the various types of gene library