Development and testing of a DNA macroarray to assess nitrogenase (nifH) gene diversity.

ABSTRACT: A DNA macroarray was developed and evaluated for its potential to distinguish variants of the dinitrogenase reductase (nifH) gene. Diverse nifH gene fragments amplified from a clone library were spotted onto nylon membranes. Amplified, biotinylated nifH fragments from individual clones or a natural picoplankton community were hybridized to the array and detected by chemiluminescence. A hybridization test with six individual targets mixed in equal proportions resulted in comparable relative signal intensities for the corresponding probes (standard deviation, 14%). When the targets were mixed in unequal concentrations, there was a predictable, but nonlinear, relationship between target concentration and relative signal intensity. Results implied a detection limit of roughly 13 pg of target m

SUBMITTER: Steward GF 

PROVIDER: S-EPMC368376 | biostudies-literature | 2004 Mar

REPOSITORIES: biostudies-literature